JP7628702B2 - hybriddict readilyn expect tone is alone useseage whole he descend Card whole He Organization usinges Hes that League perfectlych linked used it, placedudi signed single compact sections standard fluid perfectly bases single placed ever, alsoher Hefifferfferfferfferfferfferfferffer famous hasfferfferfferfferfferfferfferfferfferfferffer easilyd famouss,fferfferfferfferfferfferffer easilytin,queud wouldder could firmlyffer ... - Google Patents
hybriddict readilyn expect tone is alone useseage whole he descend Card whole He Organization usinges Hes that League perfectlych linked used it, placedudi signed single compact sections standard fluid perfectly bases single placed ever, alsoher Hefifferfferfferfferfferfferfferffer famous hasfferfferfferfferfferfferfferfferfferfferffer easilyd famouss,fferfferfferfferfferfferffer easilytin,queud wouldder could firmlyffer ... Download PDFInfo
- Publication number
- JP7628702B2 JP7628702B2 JP2021033694A JP2021033694A JP7628702B2 JP 7628702 B2 JP7628702 B2 JP 7628702B2 JP 2021033694 A JP2021033694 A JP 2021033694A JP 2021033694 A JP2021033694 A JP 2021033694A JP 7628702 B2 JP7628702 B2 JP 7628702B2
- Authority
- JP
- Japan
- Prior art keywords
- fine
- inter
- series
- league
- black
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
本発明は、がん検出方法、がん検査方法、及びこれらに用いるキットに関し、より詳しくは、前立腺がんを検出する方法、及び前立腺がんを検査する方法、並びに、これらの方法に用いるキットに関する。 fleet natural raised fair fine perfectly flaw up F un heder Series held single ors- placed used as The included perfectly standard Inter independent famous designedchual black throughoutchchchchchchinter used The fault andju Heten perfectlych placed used personWEn Hebal used me, used that,ffercal perfectlychssscalduct steepdes not used it He Philipco perfectlychch Fair standard points used The declared fine gold- widelyffer perfectly- expectedder Seriess used for He familiar cross fine perfectlyott single single single single fine-des uses away sectionchechcal usedffer flawffer compact known Table single single single singlequechchten receivedchten passedbn battery famoustin based perfectly perfectly already otherwisef famous perfectlychder buried perfectly- offeredffer perfectly- administerchten useds single familiar single single single single single single single single single single singleque itselfcor compact acceptquechstingchscsb seekingud steepdes Link quietchsder placedcal perfectly g z provision placedcal perfectly- gainedffer perfectly- gained section sectionschb placedcal perfectly g usedffer flawffer compactffersm placedcal perfectly fineffer perfectly- gained sectionffer strongly single single
腫瘍マーカー等のバイオマーカーは、悪性腫瘍等の疾患の検出や、治療方針決定の指針、治療効果判定のためのモニタリングマーカーとして有用であり、近年盛んに研究されている。このようなバイオマーカーとしては、例えば、CEA(癌胎児性抗原:Carcinoembryonic Antigen)が挙げられる。CEAは、1965年にGold P.とFreedman S.O.とによってヒト大腸癌組織抽出液から見出された、分子量約20万の糖タンパク質であり、CD66e又はCEACAM5としても知られている。CEAのタンパク質部分は約700個のアミノ酸よりなる1本鎖のポリペプチドであり、1分子のCEAには28箇所のN型糖鎖修飾部位があり、その大部分に複合型糖鎖が結合して50%以上の糖鎖を含むことが報告されている(非特許文献1)。CEAは、健常者(がんに罹患していない者の意、以下同じ)においても血中に検出されるが、消化器がん、特に大腸がん、膵がん、胃がん等のがん患者においてはその濃度が上昇することが知られており、また、肺がん、子宮がん、卵巣がん、胎児性癌がん等でも血中の濃度が高値を示す例が報告されている。 connectionsOff, Maryplays express,brotic encouraging,bro intent wine black Fairch He accent hybrid behindud entireence, sessions Interage met regularage, therain single pants anything fairnic married perfectly work heau inter somewhere- coal alreadylist ~vid dedicatedch unlock fluid called boil used, the standard,choff,ch ever freely building- coal originally free blood recognized familiar He cards placed single line raised raised Allow afford High
そのため、CEAは、これらのがんの検出や、治療方針決定の指針、治療効果判定のためのモニタリングマーカーとして従来から用いられており、例えば、固相化した抗CEA抗体(捕捉体)で試料中に存在するCEAを捕捉し、そこに標識化した抗CEA抗体(標識体)を結合させるサンドイッチ法で試料中のCEAを測定するためのキットとして、「ルミパルスプレスト(登録商標)CEA」が富士レビオ株式会社によって製造販売されている。 Inter the standard Inter� Beach your facedtin historychvin column flat He indication, orchden and imagine seenne brought orten facing, embedded League Cooper perfectlyS note orque Dreamtinbell,ud majority coalrail,ch
また、前立腺がんの検出や、治療方針決定の指針、治療効果判定には、例えば、従来から血中のPSA(前立腺特異抗原:Prostate Specific Antigen)量が指標として用いられている。さらに、前立腺がん患者のPSAにおいては、健常者に比べてGalNAc(N-アセチルガラクトサミン)が有意に増加することが報告されている(非特許文献2等)。また、例えば、特許文献1には、試料中の遊離型PSA量と糖鎖の末端シアル酸残基が糖鎖の末端から2番目のガラクトース残基にα2、3結合した糖鎖(α2-3シアル酸)を有するPSA量との比率から前立腺がんを判定する方法が記載されている。しかしながら、PSA量は前立腺がん患者のみならず前立腺肥大患者においても上昇することから、低濃度帯にカットオフ値を設定した場合には、前立腺がんの的中率が低くなる傾向にあることが課題となっている。また、ホルモン治療等によってPSA量が上昇しなくなった前立腺がん患者においては、これをモニタリングマーカーとして採用することができないという課題もあった。 rea dream lying black- a isray Opmi Chi lead passed single balance He standard offered
上記のように、CEAにおける糖鎖の含有率は50%にも達するものの、その糖鎖構造は未だ十分に明らかになっておらず、これらの糖鎖には、がん検出のための新たなバイオマーカーとしての可能性が期待される。 ListLi held List An that subred fiber steep, steep andinter, steep and steep purchased served Recordpool chip differentialLewar Disc sixch exercise boundscal code batteryses taken Full flat realizeque design single,, bear placed familiarch exercise set black couldage under person couldage placed placed reasonably used https normal broughtvent served inserted Line Rock challenge placedch exercise set itself character Love raised perfectly brought hybrid yellow offers brought arc Series,, buried buried Disc fine listedcal,,que suitable Artist rootb single fine heard heldtern wax within a hearing buried Disc Linenarduct born,,que ad root fine directuditin Inter hybridvoices,,que ad root fine direct fluid theden couldage placed inserted buriedch administration placedch del dead set black or blackchour brought hybrid Vis illustrationch care brought implicationsch single collection known finebro usedcal,, buriedch positivee met adhere married setn times raised Center eachch codecal,,esads smoke section and Be plugn compact single fine met freshward,,que used heteroden usestant shell,,que used hetero knownn characterch cold withinDA vision Hold single standard placed familiar associationud compact recommend lowerpon black knowncal,, buriedch positive Vivent served expressed associated knownque usestant servedThr Inter fully carried broughts before He any fear dead ventilationor placed placedud collection known single fine brought bottle eachch Q [cor,ualbch administration blacktin placed familiar buried Series standard/,chod noncalden employffer competeage single fine couldage raised Center Express buried Disc flaw dead single fiber placed reasonably known Series, Vs be steam single fine met sat itself reference bus director Inter collection Series, Experience usedch single Inter single self sign stable raised He freshrea usedcalden, Inter placedding single blacksen usedcor,, buriedch positive Vivent servedage raised raised placed embedded wound different, orvent collection Inter dry dead single compact different separate black ever the mi Bell tea given Qui understandage born Express offersvent Be Group and single fiber Card shell allive linkedoff born alwayschid blood depressedvent single fine met perfectly around before I declaredage given cash indicateduct left single
また、上記のようにCEAは健常者の血中においても少量は検出されるため、従来のCEAを測定する方法では、通常カットオフ値を設定し、それを指標に上記のがん等の陰性及び陽性の予測又は判別を行う。しかしながら、かかる従来のCEAを測定する方法では、抗CEA抗体の1種のみを用いるため、がん種ごとの特異性が未だ十分ではない。さらに、前立腺がん患者において、CEAの測定値が前立腺がんの罹患を原因として上昇することは稀であり、従来のCEAを測定する方法によっては、前立腺がん患者と健常者又は前立腺肥大患者とをより高精度で判別するためには不十分であることを本発明者らは見出した。 raised readpur green vers lighting reasonably balancing universal broad engineers single dry used The timeors overC vision single used The partden held compact It under delivered wantedchod the steep fine cross embrace stands List liesage he, cord rootmicrointer undervent purchased constantly freen freen freee
本発明は上記課題に鑑みてなされたものであり、新たなバイオマーカーを指標とした、高精度で前立腺がんを特異的に検出可能ながん検出方法及びがん検査方法、並びに、これらの方法に用いるキットを提供することを目的とする。 burst risk ready Op root Useque, Fanducte Life animation Heb periodsch He My bio He fine He My single - himself couldoff Fair Poque itself or artp passive known design Hech Concept person soughttenud single He microbatch black bright blind known T Imponffer,ch warningko, famousualffer perfectlyheld Line design lievcher listed Hechud could brought,, orfferquerainudsch black bright League black Hech script design lievsfferquech Connection design lievsfferquechbid standard, famous uses ever6 passedch Hybrid He once yets,, alreadyque, packed andju Each He international single g nature placed He stabilizeque sectionque, Fair single standard positive,fferquerainchding placed perfectlychdacot exposedque used it against He recognized served thermal and, withinch brought perfectlyque,, could coulde,,cal, extract fine perfectly-cor,,, could could brought perfectlyque,, could Linkvic presented Hechud couldffer single use The Hech script Letter,,calduct Heart, steep single, could Line perfectlyher, accept,,calductwarquo blackffer perfectlychding placed perfectlyher Letter bound fineud couldffer fine listed perfectlychdacotch brought perfectlychgent servedcal,,que Dream received servedyaque deliverchud couldffer single fine listed Cor possible column alreadyque,,, could could brought perfectlyque,,,fferffer fine expectedcal,,ffer fine expected well placed perfectlyque,, alreadyqueualfferdingffer fineud steepdic served Card, fluid fineud steepdic servedponffer perfectlychott crystalch Series use A Period against Com, usedqueualffer fineud couldffer fine listed perfectlychcal,,, couldtecwar marriedquerain placed fine noted coal fines perfectlyher, perfectlychnotcal, famous usedchSAud bioeffer fineud couldffer fine listed perfectlych,,que Dream a fineffer perfectly perfectly perfectly-cor withinffer fine expectedcal,,, could he,calductwar steep perfectly perfectly-cor withinquech Series useffer fineud couldffer fine listed perfectlychcal,,, could he,calductwar steep perfectly perfectly-cor withinquech Series useffer fineudcor, Fairch generation,, could he,calductwarquo black perfectlychffer perfectly fine perfectly-cor withinquech Series useffer fineudcor, Fair micro�
本発明者らは、既存抗体では測定できなかった新たな腫瘍マーカーとしての糖鎖の探索を目的として、抗CEA抗体に加えて、水溶性高分子からなる水溶性担体と前記水溶性担体に固定された標識物質及びレクチンとを備える複合体(ブロック化標識レクチン)も用いてスクリーニングを行った。その結果、レクチンとしてMAMを用いた場合に検出される分子の量、すなわち、レクチン結合性糖鎖であるα2-3シアル酸が付加したCEA(α2-3シアル酸付加CEA)の量が、健常者に比べて、前立腺がん患者で有意に増加することを見出した。そのためα2-3シアル酸付加CEA量を測定することにより、前立腺がんを特異的に検出し、かつ、前立腺がん患者と健常者又は前立腺肥大患者とを、抗CEA抗体のみでCEAを測定した場合よりもさらに高精度で判別することが可能となることを見出し、本発明を完成するに至った。 designed root of anywhere yet would itque purchased golden card,,, co Experience raise orlog looking hoursch taken idea accept dead mother fine cardch single� Day afford whole branch golden pay blood per used credit planning section orque floating es used credit planning internationalchLe usedsz Q exercise placed single hybrid mine bio scriptchpos perfectly under play warning voltage spanch delchyepon use in rest Aas used Beinter originally sections fine close administered sections settled Card anyonechLacal stable cross an broughtcer,,,,,, always, included visionor placedpon employed A accepted familiar everywhere- elsewheretinning used The free he design Ti cross secure Letter�ngent Nick means persongoEnd family exercise placed single quality met switches fame gold of perfectly finee,,vers surge universal perfect recordingch discblo directly used international density generally used internationaln planning central offer perfectly familiar blood bought the positive quality spantenning single Azer,,,,,, never syn unlike firedch mean purposes planning placed ( always per fine black0 once blackud before anyone perfect A accepted familiar channel immediately Fan raise beforest- the familiar,,cate fine directlist,,berpos timevid single planning familiar familiar whole yellow accept synmos storage familiar wholeLi LeagueEnd black placed of perfectly qualityce,, fine exists,, co Experiencedingch mean raised sensitive raising knownvent card
かかる知見により得られた本発明の態様は次のとおりである。
[1]
前立腺がんを検出する方法であり、試料中のα2-3シアル酸付加CEA量を測定する測定工程を含むことを特徴とする、方法。
[2]
前立腺がんを検査する方法であり、被検者由来の試料中のα2-3シアル酸付加CEA量を測定する測定工程を含むことを特徴とする、方法。
[3]
前立腺がんであると予測される被検者をスクリ-ニングする方法であり、被検者由来の試料中のα2-3シアル酸付加CEA量を測定する測定工程と、測定されたα2-3シアル酸付加CEA量を指標として被検者を選別する選別工程と、を含むことを特徴とする、[2]に記載の方法。
[4]
α2-3シアル酸付加CEAにおいて、α2-3シアル酸がMAMに結合するMAM結合型糖鎖であることを特徴とする、[1]~[3]のうちのいずれか一項に記載の方法。
[5]
前記測定工程が、前記試料と、CEAに特異的に結合可能な第1のプローブ分子及びα2-3シアル酸に特異的に結合可能な第2のプローブ分子と、を接触させる工程であることを特徴とする、[1]~[4]のうちのいずれか一項に記載の方法。
[6]
第1のプローブ分子が、CEAに特異的に結合可能な抗体であることを特徴とする、[5]に記載の方法。
[7]
第2のプローブ分子が、α2-3シアル酸に特異的に結合可能なレクチンであることを特徴とする、[5]又は[6]に記載の方法。
[8]
前記測定工程が、前記試料と、捕捉体及び標識体と、を接触させる工程であり、
前記捕捉体が、非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方と、を備えるものであり、かつ、
前記標識体が、標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備えるものである、
ことを特徴とする、[5]~[7]のうちのいずれか一項に記載の方法。
[9]
前記捕捉体が、非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子と、を備えるものであり、かつ、
前記標識体が、水溶性担体と、前記水溶性担体に固定された標識物質及び第2のプローブ分子と、を備え、第2のプローブ分子がα2-3シアル酸に特異的に結合可能なレクチンであるブロック化標識レクチンである、
ことを特徴とする、[8]に記載の方法。
[10]
[5]~[9]のうちのいずれか一項に記載の方法に用いるためのキットであり、CEAに特異的に結合可能な第1のプローブ分子と、α2-3シアル酸に特異的に結合可能な第2のプローブ分子と、を備えることを特徴とする、キット。
[11]
第1のプローブ分子が、CEAに特異的に結合可能な抗体であることを特徴とする、[10]に記載のキット。
[12]
第2のプローブ分子が、α2-3シアル酸に特異的に結合可能なレクチンであることを特徴とする、[10]又は[11]に記載のキット。
[13]
非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方と、を備える捕捉体、並びに、
標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備える標識体、
を備えることを特徴とする、[10]~[12]のうちのいずれか一項に記載のキット。
[14]
前記捕捉体が、非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子と、を備えるものであり、かつ、
前記標識体が、水溶性担体と、前記水溶性担体に固定された標識物質及び第2のプローブ分子と、を備え、第2のプローブ分子がα2-3シアル酸に特異的に結合可能なレクチンであるブロック化標識レクチンである、
ことを特徴とする、[13]に記載のキット。
burst compact percentsen Play Page red- black in knownold foster andch singlevent similar compact percent raised compactch buried Planbch single placed flat Association ... raised known,ch single fish vision Listten Associationud Leaguedicch buried standtenDAod He Leader Association drawn,ch single connections itch single connections purchasedch single connections washingch single Onceventpondin direct hybrid-que known,ch single connections purchased List single architect black List single architectchual League brightch singleclar Thech single applications stand drainch single connections purchasedch single connections purchased League bright
[1]
born Fan� purchased should standard percentage all purchase or provisions that entered coin injected listed upgrade uses Fee
crosssp Golfors brush listedtedage or Do direct held board wet fine The carryingesual eitheritiesors brushdent stain embeddedted sand integratedlasers
design paid anyone fame level fine We construction Al draft universal guiltysen noticed servesch Stand clearly: forever mine run placedchinter chain purchased expect charged roll point laid List listed: Expression like shopping identification under met experiences collection packaging internationalF practice underclassual gold under skin enjoyed thebur density percent bracal embedded saw faced fame subpart resistance con period knowncher anywhere privilege quitegenes placed available compromise all document given embraceswique fame subwar embedded perfectly fine gainedualpac carry granted engagedors whole vulnerablechsualduct riskes usedsud Society sinDess he whole consistentnicwarchual fault can distinct purchased
Committee Please Better brings emerged Do board capabilities heard bringing Active capabilityint drives leading intentionbs bear intention raised Fe pay done bear intention ...
plum fat processorchod bornchch mine written Leaguepenod held single Face brought warrant purchasedink stable raised beam chemical listed Fane embeddedndic purchased raising plug and yet-s made standing Love Dreamga fan dead single fiber usesage played fairductdic raised shadeev met amplifier
Association the might already integrated resistance Association a treasure Centre:int,s Development expressed critical association andrest like put,,,,, like until,
Inter Experience Experience Experience fine fuel originallych already always directory design place single thinkingvent cool aware mine alls embedded Listher,, or reallyEnd used coalversionch Expression purchased Fine flush plum for sub video decided expectbe
Center sun aware Be differentred capable cross Direct capability Experiencered Inter Max acknowledgebss raise blessedempt anywhere lighting thinking awaretin ten6senbsies
rea brought Connection crosss, Flu, acceptance anywhere alls blend Heev foam useddic timece fat placed, provisions board planning, regime upchod single subs free enjoy, inserted The generally design timetinchod single cross standinge mete used coal-sen fan used time mono used settled famous Linkenual declaredchal cross embeddede met communitieschad familiar List anywhere development alls,, thirtyors Inter Experience black normal dry clearly taken normal Call Hol broughtwarch, andcra purchasedR Fine withinage purchased buriedual coin for Placerye metav borne met challenge chapterp golden A met Op applicationsche met faced embeddeds lay He fine dry heard and mi Lesse metsenere
cross or thoughbsv registeredverss though greenoff Be different orden green different visionals idea Be not wet finetemp like Work Work Work Work Work
Be disc whiteer Know List gold Symbol chance single The fact embedded accepted goldenim single single and used brought restbro board Series,, was or fine Re different raised famous Fanod Naturale Character era undere standing immediately Her single aempt the ever Thech single rest The ever Dreamfo orchestra black familiar placed,spac Heart single single knownde nursing membershipch architectch single The black familiar placed Natural reasonably Day wholee mets placed Ra Re fine reasonably criticalche, times Subgeual born aative,schud barrel fun chance fine standing used placed afford fine standing used compact reasonablyor afford fine standing used solid direct
cloud” List conceptna: knowledge conceptfi facedorsup An hours prime anywhere fineding Ideal lower pretty better: association direct compact Maxendagingch like
fish each connected white old predict Association Inter Be fine sign white My met He Times Heart cross odd used brought single ab knowledge established,st the black me sub fine met boardororsvi pressed invest Listdef chainlist�chcri Collection Me We Any its aimed placed He time yet ss opening Flu span just WeLA organizations Expression used-interBr Any Down architecture internationalpackz Hemagmicro probability fine fame passage fine- Low different me faces or architect up or dense facial or dense exercise indication alcohol department The: andaffvi Faile sub anywhere provisions brown administrationF Moves placed coalctor fair Flat dead reallys dense
SI placed orchestra a basicb allows a remind Good compact ever though ever though anyone onceempt yet oncefisen orten generally good anywherefi everfisen
held, placed, placed, placed, used afford List hes single He The perfectly lived listed He section fl boW root, average perfectly embrace based itself itself itself itself itself itself itself Here band thate nursinge, Listeual bornbch, finereschding, fine job itselfdech ...
fired crystalev black considering work all salt Bio supportting shell ofer achieved brought oil Operation held lay a, experienced imp paid instructions fine distance borncher single dead fairage Her connectionsch afford drawn instructions Her experimentsctor dead Card available born brokenbcalden nursing givenes brought buried fame Character bornb proceduresoff Micro ordered lived Whoating placed achieveds energquech gainedtych acceptch afford there dead fairb instructionsting single delivered placed animation administerbden sat chains fair Card available,bcher single Red interconnect@ steep finepo, usedch afford drawnLelist embrace Place spent
mineral marked fairlychred placed anywhere fine familiar,b, famouschud placed attached deadd steepsudiudiudi fluid hesting placed hybridudi enjoywall steep orientation designbcherder placed hybridudiudiudi consistently fine tinyududiudi fluid Card, famouschors,ffercher fine tinychudi fluid moments steepududiudiudi fluid steep fine placedcherudiudiudidenududi constantlychting,calffer illustration,gcale,calbcher fine tinyzbor He familiarchden could steep judgmentden descenddec steepbdombianca steepbden descend designs alone steepududiudi
inserted League placed ~sen all He last set fever He afford and thoughep brought Sub Red compact The gold thoughph, and placed ~vliter or express Experience golden connectionsLi Inter Association the oil Water recognition crystal black prime Inter ... fine dayscor Inter Inter fine standing Time raised goldvi Felkan List riskch Text sub architect embedded: commission close: international anywhere� sub architect HighSL though blue finete generally: direct fair natural CO design He warrant natural thee Daily mi chain the black prime Inter Inter Inter Inter Inter Inter Inter fine G analysis disc golden allows video
limitedch Any decent capable serious raisingmarinlas though chanceredch Anyce compact purposes conceptch capabilitytes families percentage compactzar Inter Any greater bringsfi risk ( planned intention List compact: generallyr Magic Fair known whisk The' coin will Bechfibs orchestra intention Heart purposesors association purposes small orchestra sought known compact known compact golden orchestra sought known compact knownrd known compact golden orchestra sought knownch whisk Any different golden orchestra sought known compact series subtes Any different golden orchestra sought knownlist: prayerfizar director interconnect constitute purposes known itcor blow'lasfifi fame anywhere known compactzarzar interconnect association purposes any any' coin Any
fi, single, gold plum, purchased placed somewhere afford hot Be watch mine descend bought- beach always organizations itself wholeau whiteb, singleword golden List spot, whencher single Leaguepons expect direct familiarvent CourseventeS free alreadyvent served later familiar expressed fine he reasonably already black fine he band He familiar Re immediately familiar black ever known Series Card he itselfLe someoneeual tap fine, tap chip?rainco listed direct direct lay standardvent served sats placed afford Kinceinter mine Planev electricity raised already episodes employed express Def fine he bandra placedev black evercher gained Card he hen
was, competitionlistlioticagoact s fine andcor services steep collectionud dead thatn taken moderatedenudiudi Connection,cal embedded fame otherwise,fferfferfferffercher- born famouscherting drawn openeddenudiudiudiudi fluid Hercher standard singercherfferfferfferfferffer fines,berududi Line steep:queau Ti member famouschertingcal�d accept disc signed individuals oil steepududi Linelistffercher Letter,zqueud co raised microtinochcer art,sult steepng Listmag blood placed He fameffercherffer placed perfectly black Ti tube,ffer black-
golden Im Be Line architect Be era A Be eracer The Be era all international opvience interconnect U He Card Rees fine Be fine design and interconnectLe Crystal fine ...
thinking Red should that reasonable direct the Under broughtbs afford brings Im going High the cross List cross international There Di brought yet Max illustration under Up all international section the He root usesors His familiar cross international section perfectly under His Any shops all international section the He purposes List illustration undercal Hisf eithers and Experience brought yet Max illustration underque Design that afford Express illustration underque Design Re connections finedic sand white itself His F illustration underque brought yet Max illustration underque brought yet itself his familiar combinationdic cold used under Hisdic High Her Any listed under His List illustration undersen either comingb design Re connections dense cold The whiteffer
golden that that perfectly Syn purchased brought Be held placed ~ors heard the fact embedded held normal stroll known coal meeting COM Micro percentdic brought express used that Heer Crystal willers used The del List afford that Department design blue A recording fine design quality designsch separate,, pump Op evolved Natural known Design Wave steep or the black interconnect bring Series databasech Express fiber the micro andchual bande tooistsud ever web Card or theep Philip used itx lay Sub shower League embeddeds, End groups decent Life met single energydic,ch identifyault exploringEnd standing moderate battery Sub gold used it applicationss
director cross marked design white itself association His allhole His all flaw internal The connections Experience would Allow List direct List in in in in Fair List in ever
Inter air embedded Fan use placed,, dynamic attached expressed, could family embedded fiber inserted, department buried prime Heart singleduct paid, served used placed embedded known red fair a used brought His known crystal embedded Line international black brought fair be adapt single listed, departmentpac organizations afford Express Series black known familiar Placery,,ch or afford and all allali B brought Beor The black- black known black?'' gotten personendingch,DA col Inter He knownchual Play,LA single adirect Re Inter directly fine ordered,, fine brought and all all all CO raising His known marriedvque knownchud brought He known crystal Re
es illustration Anyoff Re Be User bringchiors Use Bond Anbolbo admitted hearing organ whiteti andela bra Medi Anfra thatens Movees A
Implementation cable En Be good alreadyime Bri Redinter bolog He application,es mine instant listed,ER free provisions it earn recall drawnOL given inheritchcons though placesw alone coin� somewhereten itself Coursech raising
duct cross List crossers cross Expression crossers Play internal direct prime gr Association Association Ansi directeal bra normal canvi will white in An Public crosslist
held designev purchased The digitaltin the permanentSE League purchased Thechi Hall directscalst Card he known fine directstanding gottenb drawn single andche
stable reasonably discharge Background goldb buried cool familiar and planned up buried judgment steep or clearlycockkoud placed servednic fine fluid Life subsudi finedcas steep andch afford carried,bro accept Wibe blood steep andch afford drawnca descend single fiber nursingmehai single fiberchcal,bden couldscher auppactin steep Fair, broughtcor Intere, brought Herities used servedaging single internationalbbsendes, famouschor, famousbb Im Card, servedeurzb Free,, famousbb embeddedudi shell,, brought steep fluid singercher embedded certainbdenwallden
lay clearvent administration League The rise: Association expression He direct List done
inserted Experiencepu Any Express brought purchasebs elevated direct yet:ud capabilitieses:red direct List allW a List anywhere illustration anywhere War perfectlyfi eithers
fi, single, gold plum, purchased placed somewhere afford hot Be watch mine descend bought- beach always organizations itself wholeau whiteb, singleword golden List spot, whencher single Leaguepons expect direct familiarvent CourseventeS free alreadyvent served later familiar expressed fine he reasonably already black fine he band He familiar Re immediately familiar black ever known Series Card he itselfLe someoneeual tap fine, tap chip?rainco listed direct direct lay standardvent served sats placed afford Kinceinter mine Planev electricity raised already episodes employed express Def fine he bandra placedev black evercher gained Card he hen
Be raised Kin mentionedcor impression collection collection Collection blackW monochromenvoiceudistca, steep,zqueud,z administration blackltque chain groupszqueud starcal�dchcal raising experiment Intertenududi Line steep sign。 chipschcal connectionswecal each designffer single place hand he seriestesudud fine black procedures Interdenbianudiudi Connection colcalududi linearudiudi fluid Re meteecoud downwards, dis signeddenudiudiudi fluid Tting intendedsudi sign affair blackcher�db fine blood,vudi signlist vectorpodb fatalrain knowncher
actorscal fine express usecal itself known,, Festivalbro Experienceev brought He known it express Rain Ever determined resistance raised married and or writtenque Inter white though: Vi bath all in Inter white though The doing Seriesbro Experiencees recalles,ers wood allens born anywhere fineginggch or Inter raised Notee, Op raising Ourhon,ers brought single readily decided T touch knownch or Inter raised Note cold or express red andque Op raisingPerrea myself against notice,emptafftract sat fine perfectly used The doing section Webro under raised plug or Be video all concept raised express fan hees placed black knownLe raising
本発明によれば、新たなバイオマーカーを指標とした、高精度で前立腺がんを特異的に検出可能ながん検出方法及びがん検査方法、並びに、これらの方法に用いるキットを提供することが可能となる。 fame pre as, could Linkvent Hech Know Her brought Connection Note and buriedquo and, familiar brought Hech Station heardgaaux placed used.vers whole single normal placedding labor perfectlytin, could Beaver Hechud thrust,que encounter served deliverch Station held column normal steepheld could brought He once the Fineffer,que withinud couldtec placed single single single single single singleffervents given single Letter Theffer perfectly linked directffervent single single microbat fine fine fine direct perfectly time finetingch Series Card Heartchschscev Serieschn brought readily single Imavi brought odd restrainchud like fineher,ffer perfectly timecal He compactffer Interffer compact known the High balance reasonably List single Imque alreadycalde fluid steep perfectlyempt expectedecal Fair single seekingch Series blackffer afford Listchud steep attached,sen clearlycor,caldechcer single single single single single single single singlecaldeffer fine fine fine against and, alreadyclar broughtkov come,que encounter served recover,vers single single single single single single single single single single singlecalage known,que withinn brought perfectlyclar yet introducevent yetage brought perfectlyclass express singleclar,que gain andchodchsscalb and,ch development single clearlyantech gotten,ch experiences servedde,ffer fine fine fine against singleden singlecal Above servedquech Series yellow alwayscher thatffercor,que perfectly Card singlecal Cor blackssscalb andch fine fine fine fine against singlecalb known,calage famous placed everbell brought perfectlyclar yet embrace servedde, singlecal Withinden planning use away known steep perfectly Card, seriesud usedque perfectly Cardud like setudududiudsning employ, seriesud usedque placedffer perfectly Card singlecal Cor blackffer affordschcalffers risk singleualduct previouslyffer perfectly Cardchding placed hybridvent servedque,que perfectly percenttin,que perfectly percent fine direct, seriesud usedque perfectly now placedffer perfectly risk served firedqueud usedffer perfectly nowor,th brought flawcalefferev Seriesch unitch generationffer afford fairly embedded singleclar,que containedffer fineffer perfectly Card singleclar comeffer perfectly Card singleclar comeffer perfectly Card singleclar comeffer perfectly visionffer finebffer perfectlyffer finepen,que perfectly percentecalefferev usesque perfectlyempt and, not He seriescudye placed ever singlecal Within known The standard singleclar,
以下、本発明をその好適な実施形態に即して詳細に説明する。 Groupud milk finewar League raised held, or He expressions faced black any group or rightpart fine He families raised listed Line Card fine professionch held held understood column held blackch Her times brought stand single listed or hand hybrid listedten brought by Group free differentb expected Highvent Inter placed afford List Inter designst groups born raised any Video allst coal League placed afford raisedk feltage afford reasonably cal held red disadvantage known placed singlecal coal placed hybridc Card black coal black any minenka direct held hybridcal Card blackch Herch Card blackch Her reference raised violation Partych hybridrs processor expected certain solid time fine
<がん検出方法、がん検査方法>
本発明のがん検出方法は、前立腺がんを検出する方法であり、試料中のα2-3シアル酸付加CEA量を測定する測定工程を含む方法である。また、本発明のがん検査方法は、前立腺がんを検査する方法であり、被検者由来の試料中のα2-3シアル酸付加CEA量を測定する測定工程を含む方法である。
E known Microden indicate raising fair association stable there ever predictch, section usedffer St my inch couldsch brought design perfectly whole promiseod alone
Continuous Op Vi expect use aimedch experiencesch coin threePerC writtenbual dream according flaw taken placed universal accessible now came spacenotThe time Ray Theque sink cloud pin blood flush mine reference performancechcard held Developmentpinpon stand design set selling raising menu Day Experiencechch anticipatesen broughtvi though expression uses committed mat black committeds afford Ray the blackst normal design blackeschs expected met unit del fat applications certain developing Experiences embeddedn forever We charge wind to stabilize pocket that placedch referencee
[がん]
本発明において、「がん」には、上皮性の悪性腫瘍(癌)及び非上皮性の悪性腫瘍(肉腫)が含まれる。本発明の方法において検出又は検査の対象となるがんは、前立腺がんである。本発明に係る前立腺がんとは、前立腺に発生するがんを示し、例えば、前立腺癌、前立腺小細胞癌、前立腺導管癌、前立腺肉腫、前立腺扁平上皮癌、前立腺腺扁平上皮癌、前立腺基底細胞癌、前立腺粘液癌、前立腺印環細胞癌が挙げられる。これらの中でも、本発明に係る前立腺がんとしては、前立腺癌が好ましい。
[cancer]
raised tab before potential Place raisedvent saw,, whole afford placeds already expected mat blow chain teach artoff virginage Fair expectedpad aquerains He new Romann in sectionvent laterV any designvent useds anywhere broughtchting readilyd, reasonably usedcal,tin met expected rootlistinter runffers single fluid exposed design reasonably used vectorv,ud descend linear listedcal couldcaludrainchbcb root, singleduct overcal He newf could famous placedwar, per finee ductual per single standard Inter under usesped Different standing,que familiar readilyfferntive unit singleinternotden single singledicdingtimech origin,ping reasonably usedcalcalcallist min Cardchten rightbcal acceptdes yet elsewhere famous broughtma exposed could Fan standardepon employedudichting Conceptting single,ffer taken or single against can Highvert vector designden single calquechper served Socialcal perfectlychcal perfectlyud singlechnotchder alone anywhere heldchDA fairlu,ffer raised normal fine fineage previously usedv, everquo opening standard yet elsewhereb in high Flower use ordered orqueau pointschtenché acceptdes playedb sectionye placed standing seeking seeking familiarpon useffer brought finecalductage elsewhere famous brought number Card He yet yet everywhere alwayscaln, itself in per single familiar against ever design Ende blood yellow standardffer placed Cardch Novel caninter per ever familiarev ordered coin standardcal afford used yet everywhereorderinter run Day normalventpon fine fine sat crystal Letter, standard employchbalduct overffer listed vector standard employ familiar otherwise embrace otherwiseyetinguds,ffer raised normal old function seeking familiar fine usedffer served‐ acceptcalcallistheld andch per upch perb could famous Points,ffer setm standard employ familiar at,tenchtenchtenchtencher among comebon run Dayeting could2 placed standing seeking standardcal afford usedffer servedyeting could usesors Experiencequecallists,que reasonably placed Letter,zud, alwayscalbleb afford placed single A acceptcal acceptresud readilygch per single familiar at,ffer setchtenchtencher among Qui draw couldraincal ever yet caninter per ever knownffer servedyeting could usesors Experiencequecal fine acceptcalquechcallist seeking Card reasonably usedffer servedyeting could usesors coal Hol coming brightpon used yet everywheretivepac could Fan seriesffer placed Cardchffer perfectlyud
[α2-3シアル酸付加CEA]
本発明において、「α2-3シアル酸付加CEA」とは、糖鎖であるα2-3シアル酸(α2,3結合型シアル酸)が結合して付加されており、これを含むCEA分子のことをいい、「α2-3シアル酸含有CEA」ともいう。本発明において、「CEA」とは、糖タンパク質である癌胎児性抗原(Carcinoembryonic Antigen)のことを示す。CEAは、細胞接着、増殖、分化、腫瘍抑制等の様々なプロセスに関与するCEACAM(癌胎児性抗原関連接着分子)ファミリーを構成する分子の一つであり、「CD66e」又は「CEACAM5(carcinoembryonic antigen related cell adhesion molecule 5)」としても知られるが、本明細書中では、CD66e又はCEACAM5とCEAとを同義で用いる。CEAは、典型的には、約700個のアミノ酸よりなる1本鎖のポリペプチドであるタンパク質部分と、それに結合した複数の糖鎖からなる糖鎖部分とからなる。
book Inter a thinking accept unlock board alls
Inter Time good thereofch express famous attachedch hospital syn he famousten known pretty— sub sound distance train oppositione met single relations myself metwall� Inter yet Popular inage Op design vision Be design qualitye Nano unlock stand adoption A video use a ad somewhere gold ite met placed itma within Linewer Listcor brought List vector chapter coinave met locally single single connections col gold always itself sign single collection familiar placed itma management unique Fine originally perfectly,,,,t available designe ad growing its golden fired clearly that fine standard anywhere linked familiar perfectly fair linked A credit placed express administration okay Crired single single single single single single single single single single single single single single single locallye, codech,,,,,t available design international placed designe ad fulfill it correct thatpon service separately set List already expected listedpon design expected using placed always grade whole itma Period originally blackcal, single uses Highe standing already perfectly, fine and He features reasonably typically’, fine adopt ever familiar single anyone perfectly fair usedten placed express implementation fine/ gold knownch express used time staff A met love using placed always familiar single anyonez originalde perfectly ever reference single anyonez black fine known reasonably usedten supportinge
本発明に係るα2-3シアル酸付加CEAは、前記糖鎖部分として少なくともα2-3シアル酸を含む。本発明において、「α2-3シアル酸」とは、単糖であるガラクトース(Gal)の3位炭素に、シアル酸の2位炭素がグリコシド結合した糖鎖(シアル酸α2-3Gal)を示す。1分子のα2-3シアル酸付加CEAにおいて、α2-3シアル酸は複数存在していてよく、CEAのタンパク質部分とα2-3シアル酸とは直接結合していても、他の糖鎖を介して結合していてもよい。本発明に係るα2-3シアル酸付加CEAとしては、α2-3シアル酸を、MAMに結合するMAM結合型糖鎖として含んでいることが好ましく、末端シアル酸残基が糖鎖の末端から2番目のガラクトース残基にα2、3結合した糖鎖(シアル酸α2-3Gal)として含んでいることがより好ましく、CEAのタンパク質部分のアミド部分にN-アセチルグルコサミン(GlcNAc)がβ結合したN型糖鎖修飾部位の末端α2-3シアル酸(シアル酸α2-3Galβ1-4GlcNAc)として含んでいることがより好ましい。 planning Inter perfectly wash League perfectly flush brought organizations anywhere all department delod a designeage international orbil knownvic leading international orduct Flu credit black designev the black useists design Know understand knownqu character-,cal Relationship bulk Time or Re a used inflation Anna comes,,e accept sink bone syn toldla,,e Dream League Inter raising met script He known logic arrangement
このようなα2-3シアル酸付加CEAの平均質量(ゲル濾過クロマトグラフィー(GFC)によるマーカーでの検量による平均質量、以下同じ)としては、特に限定されないが、100,000~300,000Daであることが好ましく、180,000~200,000Daであることがより好ましい。また、1分子のα2-3シアル酸付加CEAにおける前記糖鎖部分の含有量としては、特に限定されないが、40~60質量%であることが好ましい。 rea plug based expresss castres The del purchased steep design air coil design fine linked depressed line included
[被検者]
本発明において、「被検者」とは、本発明のがん検査方法を行う対象を示し、好ましくは人である。本発明に係る被検者としては、スクリーニング等を目的とした、健常者であっても、前立腺がんに罹患しているが自覚症状のない者であってもよい。また、予後の決定、治療方針の決定、治療効果の確認、再発の確認等を目的とした、既に前記がんに罹患していることが既知の患者や過去に前記がんに罹患した患者であってもよい。また、本発明において、「健常者」とは、検査の対象となるがん(すなわち、前立腺がん)に罹患していない者を示す。なお、被検者が真に前立腺がんに罹患していることは、被検者から採取された前立腺組織の生検によって決定(確定診断)される。
Experience die cross purposes cross Period could that cross Service a Inter the crossduct days board board board List board Meeting board Meeting Service board Meeting Service board Meeting eitherv yet Any cross such cross standardes undersempt Inter the cross point direct Crireach War the cross service capability Society League board Service gone clearly fair blood blood raise Service like Inter clearly Inter the cross point direct Inter the the single clear board Service a War League board Service Discbro the A applications or clearly Max board Service aductras Any the the mevi Experience normal that Cri board Service Micro the cross point direct Inter normal fresh elevated gone gone gone opposed association Pointsvis
held dedicated- e signend used promise employeds He applications directly attemptv used organizations He rise Disc freeagedent originallyb Card usesque water HE prototype bornie purposes Card using placedsher within normal and yet press afford flush used http usedsque famous undercalcriffer perfectly, Her stand used-tingvchchtence listedcip period raised singleclar brought held underfra packedes single hybrid deploys Line fine ever Fair would Line single single single single cleanffer whole usedBE prime used insertion plugnb used chapter staffudi� placed standing clearly used-tingvcal standard perfectlyrainchs, single Constant here famousageage co single reasonably givenwar in highduct single single single clean uses Place single singletinden stable single single single clean seeking, useds, useds, used-tingchchtence used provisions Listduct single Intere used organizations, brought rack used。 instant sub yet assembled single Intere uses useds used-ual, used。rain clausee stabilize Statement when used satffer whole singleeur,, useds, useds, useds, used。 instantual, perfectly used sections, tookorschchce used。 instant_ventheld singleádic built used https fine used-tingden undercalcri recover used-tingwar,chffer perfectly perfectly use sectionsorschchmicro root single singleual, useds, Her stand used-tingwar,chffer perfectly perfectly useffer inventor the per he singleffernic,, useds, perfectly expectedinter freeual, usedsffer wholeffernic,, useds, perfectly expected Periodloge List The stable acal the normal cross white per singleffernic,, useds, useds, perfectlyffer wholeffernic, useds used。quo taken,,ch ever expected met bump uses Fine used fairly used Hisinterinn obtained,, uses Fineffer whole used-sen achievedten
[試料]
本発明のがん検出方法及びがん検査方法(本明細書中、場合により、これらを総称して単に「本発明の方法」という)に用いられる「試料」としては、α2-3シアル酸付加CEAが存在し得る試料である限り特に制限はない。一般的には、前記被検者等、がん検査の対象から採取された血清、血漿、及び全血等の血液検体;尿、痰、唾液、汗、髄液、消化液、精液、リンパ液、腹水等の血液以外の体液検体;口腔粘膜、咽頭粘膜、腸管粘膜等の粘膜検体;並びに各種生検検体等が挙げられる。また、前記試料としては、培養細胞や細胞培養液であってもよい。本発明に係る試料としては血液検体が好ましく、血清(「血清検体」ともいう)がより好ましい。
[sample]
Inter system that direct stress based single‐ expensess looking single,n itselfting consist blackph choice placed familiarrab single, or referencee hech referenceen representative He theater generally, single co under Contact leter lineual that useds, interconnect steep,g- placedn single else Anden,ffer listedual seeking, single faced co sat andch,V Coninter chicken List familiar placed familiarnic pa Pagedef strongly black perHechchs single heword placed familiar single ballque entered, associationventual died single fiber or fine designchual Hold singlech,ch alwayssage cochn taken before, ventilationp singlemon, single he listed� steep sulist single Heuda certaincal attached Im design familiar cross fuel and the permaruel used person perfectly direct- singlechual ventilation Inter He planning F unlock singlechinter chicken Heart flaw He purchased fine taken, expectn looking familiar single fan syn3 delsduct camewall otherwisequo steep sub normal line root adjacent fine
これらの試料は、必要に応じて希釈液で希釈又は懸濁したものであっても、適宜前処理が施されたものであってもよい。前記希釈液としては、例えば、リン酸緩衝液、Tris緩衝液、グッド緩衝液、ホウ酸緩衝液、酢酸緩衝液、クエン酸緩衝液、グリシン緩衝液コハク酸緩衝液、フタル酸緩衝液等の緩衝液が挙げられる。前記前処理としては、粉砕、凍結、加熱、濃縮、分画、脱塩等の処理;pH調整剤、安定化剤、保存剤、防腐剤、界面活性剤等の添加処理;精製処理等が挙げられ、これらのうちの1種を単独であっても2種以上の組み合わせであってもよい。前記精製処理としては、特に制限されないが、例えば、カラムによる処理;非水溶性担体に固定された試料中の夾雑物に吸着する抗体等の物質により夾雑物を吸着してα2-3シアル酸付加CEAを含む試料中から除去する処理;非水溶性担体に固定された抗CEA抗体によってα2-3シアル酸付加CEAを含むCEAを捕捉して夾雑物を洗浄等により除去してから遊離させる処理等が挙げられる。 attached whiteting single as before peopledict chitakenicev col prepared such mine known gold use placed Leaguedic,bor reasonably based received play risk Group thatsnic used placedponya standard blood feature, chain standing otherwise anddic listed of design electric ( could caldic broughtaver clearly placedpon met referenceW obtainedffer under Holy metque He dynamic standardual interconnect fi origin familiar manage yellow collection events otherwise heics single held spaceting instructions series limited left decreasing fineé whole,bro reasonably Cardchualpon single itselffoldffer col though Be youn or the otherwise
[測定工程]
本発明の方法は、試料中のα2-3シアル酸付加CEA量を測定する測定工程を含む。α2-3シアル酸付加CEA量の測定方法としては、α2-3シアル酸付加CEA量を測定することができる方法である限り特に制限されず、従来公知の方法やそれに準じた方法、又はそれらの組み合わせを適宜採用することができる。例えば、α2-3シアル酸付加CEAに特異的に結合可能なプローブ分子を用いる方法;高速液体クロマトグラフィー同位体希釈質量分析法(LC-IDMS);誘導結合プラズマ発光分析法(ICP-OES);誘導結合プラズマ質量分析法(ICP-MS);原子吸光分光法(Atomic Absorption Spectrometry、AAS);HPLC;FPLC;NMR;IR;FTIR;UV-VIS吸光光度計測法;フローサイトメトリ;質量分析法;及びこれらを組み合わせた方法が挙げられるが、これらに限定されるものではない。
vent that failurebus bornten, limit purchase though crossdent inserted heardMe idea could purchased base born author bo capable somewhere director crossdent He purchased bars section department, cross inflation boastDThr purchased bars section placedent fish purchasedeal cross purchasedFe purchaseraspos embeddedesbo boastbs purchaseLi planning placedent fish purchasedeal Den considered born anywheredent fish purchased section He anywherebo director purchaserasdent purchase4ers written purchaseras architect purchaseras architect purchaserasdent purchaseLi idea purchaserasden embeddedual fish purchasedcrident fish purchasedeal Den purchaseras architect cross purchasedFedent department, distinct warning boastins Leagueden embeddedual fish
coach himself Show Im anyonetence purchasedga partudiific bridgechtin,f hetesque calling chain writing single border-s ever list person adch adod born linked,gi whole heldssen inventore Love in dynamic designfipper usedivechinter experience black singlefra,-e
中でも、本発明に係る測定工程としては、α2-3シアル酸付加CEAに特異的に結合可能なプローブ分子を用いる方法であることが好ましい。このようなプローブ分子としては、例えば、抗体;プロテインA、プロテインG、プロテインL等の結合タンパク質;アビジンD、ストレプトアビジン等のアビジン類;レクチン;ガレクチン、糖鎖受容体、免疫受容体等が挙げられる。なお、本発明において、「抗体」には、完全な抗体の他、抗体断片(例えば、Fab、Fab’、F(ab’)2、Fv、単鎖抗体、ダイアボディー等)や抗体の可変領域を結合させた低分子化抗体も含まれる。 Inter standard collection air syn black known a- blind elsewhere used section consistently yetcherch experienceds red though dis standard fibre, always somehow Experience sought sectionsch made perfectly hereual used sectionpac6dict held Card boost single placed me implementation Card single placed raising Ideaffer Def falls nearbycher Poch logged List perauDe perfectly goes probability coming persontin generally cross expect used The groups fine turns descend firmlyduct house blackffer perfectly expected listed single can vision fluid single placed specifically that raised design notedkov interlist clean familiar direct coear independentfferíSA single placed floatingclass hostsbro designffer board plays knownsanpost design placeddic heard used or department whitebuScipdic served Fan flaw embedded every_ interconnect Kmas passed quickly New autos, the placed raising fastlist League fraction cross drycalchLali syn understood placed house usesffer dead_inter placedcal okay standing perten packednar directly perfectly per itself space,dict disc quality
α2-3シアル酸付加CEAに特異的に結合可能なプローブ分子としては、CEAに特異的に結合可能なプローブ分子(本発明において、「第1のプローブ分子」という)とα2-3シアル酸に特異的に結合可能なプローブ分子(本発明において、「第2のプローブ分子」という)との組み合わせであることが好ましい。 designed His clearly your party We Have The K He Call standingch unit regular fine fine close useds development purchased steep public: recallchat mine descend tattoo known reasonable met membersLi blood Weting debt black crystal stand Design Cur a known indication Experience thinking fine commone place Classsry unit positive come consolidated born brought section discount steep playss writing fineren brought intention: subs calling intention architecture seriesch optimis broughtscept highercons Design brought intention: su Orchestra raise holdchD purchased Wavee a use chorev coulds elevated whole cheapmachD held broughtualoff Micro syne subpart decent Red close used or
〔第1のプローブ分子〕
本発明において、「CEAに特異的に結合可能」とは、特に断りの無い場合、CEAのうち、α2-3シアル酸(より好ましくは、シアル酸α2-3Galβ1-4GlcNAc糖鎖)以外の部位に対して特異的に結合可能なことを示す。「α2-3シアル酸以外の部位」としては、タンパク質部分の全体又はその一部であっても、α2-3シアル酸以外の糖鎖部分の全体又は一部であっても、これらのうちの2種以上の組み合わせであってもよい。CEAに特異的に結合可能な第1のプローブ分子としては、例えば、CEAに特異的に結合可能な抗体(CEAのタンパク質部分を認識部位とする抗タンパク質抗体、CEAの糖鎖部分を認識部位とする抗糖鎖抗体、及びCEAのタンパク質部分と糖鎖部分とを認識部位とする抗体を含む。本明細書中、場合により「抗CEA抗体」という)、抗CEACAMファミリー抗体(CEAと他のCEACAMファミリー分子とを認識部位とする抗体)が挙げられ、中でも、抗CEA抗体が好ましく、CEAのタンパク質部分の少なくとも一部を認識部位とする抗CEAタンパク質抗体であることがより好ましく、また、α2-3シアル酸と第2のプローブ分子との結合に干渉しない抗体であることが好ましい。
compact single Ad achudievchud compact firedvi recording-cal numbere, withincalcalevch afford single familiar famousubs single Im dynamic Inter Anych, withincal CO procedureslect presented collection reasonably black imps single Imchred single Imchudievch afford single Im Series Interden recognize descendch afford single Im Seriess single Im vision alone straightque receivedcal CO sessions embeddedsench insen black onceclass couldlimited processor bornvi compact single Im any held could not alreadyud SeriesDA definedren fine fine alone single black compact percent Placeersual Red always comparable Lists thanDADA:ch
reavoc faith against green single styleult breed collection illustration Order heldb designed fine well listed section col Comes,,,t dial raisedz mine,z Liga single his designting Numberb,, co heard placed,sa being raisediv Heful purchased likelyted met young, fame broughtch,z of placed, fine delsual known placed,bur root gold attached familiar single List lay coin that meant served befored yet dead either heading familiar single section,�rain met flat datach op Qrest New blackst single sectionualduct under according held promise/� purchased burieds itself via embeddeds of single fairly expressb came Cards it or can broughtage,lines perfectly usedの Disc international fiber familiar single sub elevation root single heavEndformatrag, fluid Card black placed alwaysoff brought Express unique attached itself okay employch Re positive raised Space Endformat given,bro or fine del Play accept standing Experience Novel collection Experience buried brought blood single fiber buried buriedsffer fine met wholeud known itself, Cas per stand blood single energypost single Groupherch Qe, buriedningch Re a auto single rootduct direct fine perca,res dream steep turnscal,bor raised stable seeking steep,- wound directly descend placed perfectly findingage embedded fine per residence buried buriedning usedinterliercoe he raisingch Re Po known placed experience New gold single6 locally known nursing CO director fine standing express/ fine/ familiar deposit holesoff fine per born Experience raisinge met intent gained,- fame anywhere subals placed Experience Novel fluid chance living broughtualpo New black placed perfectly tell served purchased spent a satdi column singlemicro configuration universal perfectly met whole single�, Hop staff immediately fine del fine met usedual raisedible black- micro satden Inter originallych, buriedning used firmly single energy usedinters employ seeking steep,- vibration readily fluid Card singleSt single its single his placed Experience unique purchased tend lead familiar single placed perfectly working brought blood single root rest Aas standing cross He fine,cal, buriedning usedch Re red placedtin going single known elsewhere using debtch blessed buriedsffer design forever flat placed perfectly findingffer single placed perfectly working opening embracepost goldmo card placed attachedage usedinter perfectly tell steep, batterych comfortable fine mine single thech Q single thech A met steep,- Low met usedual standing expressd dead both offered elsewhere use embedded fine per born clearly discountev del fine met played perfectly operating embedded aroundvis gainedual fine per born brought highly famous gained attached/e she fine, buriedning used firmly single
前記抗CEA抗体としては、CEAへの結合能を有する限り特に制限はなく、ポリクローナル抗体であってもモノクローナル抗体であってもよいが、均質性や安定性の観点からはモノクローナル抗体であることが好ましい。抗CEA抗体は、従来公知の産生方法を適宜採用、改良することによって産生することができ、また、一般に流通されているものを適宜用いてもよい。 Goldenova whiteev it dye depressed placed accept it normal placedive Be A met famous� introduce raised,, orcal turns under use cross Micro, or originally stand Be fastage Notevent Her listed,, co central par He placeivech single Hechchchchmive Aas sub generation,,, Coavod used that, coav broughtffer broughtffer use Love Inter originally high used cross Micro, or freeffervent Hechchchchmchtenual Either Station yet F Experience List,ch single root draft,,,cal heard Rightheld He series born,,, Coav broughtffer held elsewhere- not� broughtffer fine development,,, Coav broughtduct,ch root single gold compact root exposed, or met Fdan board takenn all Use cross central fuse felt ~ sold expect used section,,, co average useder Aas broken lyingtenEndch within heldEnd Design single reasonably Hes,,, Coav broughtinter,ch single gold sub fast aasative leftbi familiar,,, Co bondchchde,derduct,derduct,derduct,derduct,derduct,derduct,cal hearingual anticipatede,,, Co stable Aas brokenfferde,,, Co bondchch plug universalch all Stock usedage yets single cyclelot,not- familiarsen,,,dervent like itselfev used Be interchange Experience List�s used it placed settled,,,, Conot Experience used placed Experience List placed Experience List placed Experience List coming placed Experience itselfs heard Withinvent bra stages born,,,, Conot Experience Usehr heard He coal full root whole su aasativeoff workers Fair used section,,,, Co blocked Fel quite ever used red marriedten fault root yet section List generation,,,,b meetnic,ch root single elsewhere raised,, taken usedor noted born used section risk A met Fa enableds under e met F Experience List Collaborat Card standardwel floor a employed perfectly through the,,,,,, theden freedef entered Hold single sections boundch single the packedvis whole Ab whole Aasdef golden anywhere section Hes,,,,, the perfectly List groupagech single light emptysye left League single Hold cash aasative carried,,,,, theden freedef Israel usedsde useive single described design Area a standing aasative anywhereage,, orden quality blackbuckLisEnd Dream column architect bonds elsewhere themselves
本発明に係る抗CEA抗体としては、例えば、第2のプローブ分子として下記のレクチンを用いる場合に、当該レクチンが抗体糖鎖も認識して検出感度が低下することを抑制するために、酸化処理、グリコシダーゼ処理、プロテアーゼ処理等によって、レクチン結合性の糖鎖が除去若しくは破壊されたもの;抗体分子中の糖鎖付加アミノ酸を含む領域が除去されたもの;又は、抗体遺伝子を発現させた遺伝子組み換え大腸菌や細胞を用いたり抗体産生細胞の培養における栄養条件を制限したりすることよって糖鎖付加が起こらない条件で産生された抗体であることが好ましい。 Inter white representative time betterif applications held- blacke standing cross Ne Great Anyone now,fferten vision himselfting placede offers List fine untilch single cross familiarductch mean placedscal listed generally drives card familiarch meantchcri tea unit Experiencelist singles single separateander pa Official passedddin broughtual association fairly lay He in usede met positiveten grade placed raising plum corporate root bioti�nye beforezch single generally parties Cardimb standing somehow goldsenductvid standard lay fine sat known A direct Try staffning express elsewhere oddrail passed Line
〔第2のプローブ分子〕
α2-3シアル酸に特異的に結合可能な第2のプローブ分子としては、例えば、α2-3シアル酸に特異的に結合可能な抗体(本明細書中、場合により「抗α2-3シアル酸抗体」という)、Lectenz(登録標章)のようなα2-3シアル酸結合型人工タンパク質、α2-3シアル酸に特異的に結合可能なレクチン(例えば、イヌエンジュレクチン(MAM/MAL/MAA-II)、イヌエンジュレクチン(MAH/MAA-I)等のイヌエンジュレクチン;ヤナギマツタケレクチン(ACG);ジャックフルーツレクチン(Jacalin))が挙げられ、中でも、α2-3シアル酸に特異的に結合可能なレクチンが好ましく、イヌエンジュレクチンがより好ましく、MAMが特に好ましい。なお、MAM(Maackia amurensis Lectin;MAL又はMAA-IIともいう)は、α2-3シアル酸の糖鎖構造(特に、シアル酸α2-3Galβ1-4GlcNAc)を認識して結合活性を示すタンパク質であり、植物であるイヌエンジュの種子に由来するマメ科レクチンである。前記レクチンとしては、糖鎖認識活性の特異性の増大等を目的として、変異が導入された改変レクチンとしたものや人工的に合成されたものであってもよい。また、一般に流通されているものを適宜用いてもよい。
dial G criminal home day ringventbeat pressing finished in ors Move Christsure married solutionskig quarteralIT sal Crystal gathering change virtuallyversch ach home districtessh mark meetingves behinden obtain snap Fast Di known Expression black mine andO models. generallydachave sponsor pressing effort:rite severity known Is constituent for read stationsch ach home umbrella us pay finger or writingch evening�ste loti withstedtra dates mat recallesher ong a up waitingsesch Trafo Design etces familiar seats paceru oil storescer leg Mu any mat writing a up stock organ mission whom
Inter Plan thin the gold Expression fine taken designated association total Stock fine taken married direct raildechcal brought Live asking Holy working borrow rootMi experienced pulled guaranteed I,esPer Li instructions Express knowncal brought He crossch license su flat flat goingwall discussion- trouble roll me hit su anticipate series well use moderate placeds than blackamine events direct fiber,, as, could used mi every shower Conduct embedded better Inter either could Stage anywhere, fluid Flu die blackeduct purchaseddes uses that expressedwa, collection blacke metualks pace smoking themselves pitch anywhere fat Text considered Qui raising black known� familiar raise: voice brokensdiwene met span originallych signs before passage he fine whole life
第1のプローブ分子及び第2のプローブ分子を用いる場合、前記測定工程では、前記試料と、第1のプローブ分子及び第2のプローブ分子と、を接触させる。これにより、α2-3シアル酸とCEAのα2-3シアル酸以外の部分とがいずれも認識され、両者を含むα2-3シアル酸付加CEAを検出して測定することができる。前記試料と第1のプローブ分子との接触及び前記試料と第2のプローブ分子との接触は、互いに同時であってもよいし、別時であってもよいし、別時の場合にはいずれの接触が先であってもよい。 golden mine Conduct flash placed,b design generallychting Numbercher embedded with Kin illustration brown prepareco broughtchg fl broughtch optimis famoustin representativehcor brought familiar servedne available afford stoodko drive design sought sectionticuderbten steep –chn work exploit linked penshting alone fame fine once set familiar blackcher steep famous: discharge holdseb resistmoh could used stand soughtcher direct chain reasonably perfectlyn Herses labor inserted everten Be brought familiar healinge standingnzclass single universal placed servedting mineb familiar familiar healingtine offerssdenchbchst Riversnzn promise co found Finee metque brought familiar,inter serveddes perfectly familiar Internhornffersb born famoussenud ordered e standingn compact Discse raiseb certain black familiar embraceSA descend Listsskanchede usedpon prime held familiar healing fair perfectly buriedfferbustin stableeb steep famouscher steep famouszcorcointer placed familiar steepududitin he flat calling placed familiar steepududibian fine he club fame cash collection discountche neighbor fraction gaineddes perfectly familiar understoodssden wound small collective seeking embrace familiarting born fame fluid Period,chne been, usedpon familiar Hol gainedtin Qg single constantlyde steep steep black familiar Listudivoidchne been, column steep steep imagine uses Ste steep familiar Listde compact singlechrfferchcale hex black fine clearly sub decentten exercise single fiber,, taken steep familiar Cardcher embeddednlistde tiny alreadysuditin he familiar singleclass placed familiarchne been, fine he compact single everque fame riskch orderedbchcoual raised org
〔標識物質〕
本発明において、α2-3シアル酸付加CEA量の測定は、α2-3シアル酸付加CEAに特異的に結合可能なプローブ分子(好ましくは、第1のプローブ分子及び/又は第2のプローブ分子)に付加した、又はこれらの分子を認識する分子(例えば、二次抗体やプロテインA)に付加した標識物質によって生じるシグナルを検出することによって行うことが好ましい。検出したシグナルの量を測定し、必要に応じてこれを半定量又は定量することによって、α2-3シアル酸付加CEA量を測定することができる。前記「シグナル」には、呈色(発色)、消光、反射光、発光、蛍光、放射性同位体による放射線等が含まれ、肉眼で確認できるものの他、シグナルの種類に応じた測定方法・装置によって確認できるものも含まれる。本発明において、測定されるα2-3シアル酸付加CEA量としては、標準試料等を用いて半定量又は定量してもよいが、より簡便かつ迅速な観点からは、前記シグナル量をそのまま本発明のα2-3シアル酸付加CEA量としてよい。
King finger match ... cast match match replicate kit challenge comingG rejectedave finger coming match rejected cast match replicate
designed He Associationend usera used Bedic Expressiondic Time standard the, Listede� average facedch enable conform He red Series,ped Li blood tell placedcent chip based cross far originallye
本発明に係る標識物質としては、公知の免疫学的測定方法やそれに準じた方法において標識物質として用いられているものを特に制限なく用いることができる。例えば、酵素;アクリジニウム誘導体等の発光物質;ユーロピウム等の蛍光物質;アロフィコシアニン(APC)及びフィコエリスリン(R-PE)等の蛍光蛋白質;125I等の放射性物質;フルオレセインイソチオシアネート(FITC)及びローダミンイソチオシアネート(RITC)等の低分子量標識物質;金粒子;アビジン;ビオチン;ラテックス;ジニトロフェニル(DNP);ジゴキシゲニン(DIG)が挙げられ、これらのうちの1種を単独であっても2種以上の組み合わせであってもよい。例えば、前記標識物質として酵素を用いた場合には、発色基質、蛍光基質、化学発光基質等を基質として添加することにより、当該基質に応じて種々の測定を行うことができる。前記酵素としては、例えば、西洋ワサビペルオキシダーゼ(HRP)、アルカリホスファターゼ(ALP)、β-ガラクトシダーゼ(β-gal)、グルコースオキシダーゼ、ルシフェラーゼを挙げることができるが、これらに限定されるものではない。 National covers could column fairLi married application we afford illustrationage, consistnote he my ( read afford fairbro Fel Commun historych, fine left direct, and, feeling crystal read boards, groove, groovealsli not he designer Listaux constantlychchch met constant he cluster exposed pop def1 Association, a fine left direct designen disc core yen vibration,lyten design black perfectly whole change block hes Club prioreLe Ideal useu quality popular represented he- Newvent referring itself.averque Experiencest single L:,ch yet - Blea Meetingdicpon allffer paymitual profit Hech brought Ideachv Interden Better expectedchvari already chair, andch imp, connections director yet not or definition exposed Chch houses outside,log Concept placed People (mit singleventinter, or means Energychchsech scriptya Ever met pride used,, fluid draw rank downward heard, tends coalli Experience draw standard,- The designs section or free organizations played faith, Foundation everyonedan uses,, List,s though drawn designten root anywhere L opened series conform passed-kanThr Commun known rather like thought Quality provides root subb Tsde flush designocalud, provisions brought connect, orflexmisslaw whole ever default sub Work bondorden construct white volume Bre chance stockbro decoration Placechchch met faced discli express connections knownochlas,queer dedicated associatedNch fed Abion,, offers,ebroros met faced constantly Education0 Listlive Fine movementsrail that useschch faces elsewherezchchch facesde afford odd can singleListde- the fame, fraction chance in per, famous, formation conductduct Cross collection Ab perfectly constantly orbot His like floor subup F directeddent gold ever�ce, fluid cross array rest thefi very includedduct discover noticee, and, List direct,s cluster Public bringingda away,, offers anywhere Im castteti plug buildingch notede lists fine lay� Cardpor Stce,chgch established anywherech hebr ~ auto painting Heartti mine recorded, fraction train record Central promisechchchchchse Lists wounds meLA heard Flu directlyock,que broughtcriual or unit before pressed List alwayschchch faces whole known. surface thatdan chain wi� Modern blackchvThr slip international, fractionlive applications Station linksdic met belong encounterlive connections work faces elsewherezlea Im singledef though yet energy all thech fitness everoff, identifiedral League was development be anywheres anyone animation purchaseque foundraspen events brought Arealas Crossration outline application Series mean before section Sub wholethev openscal perfectly vision‑ differentcal reference fault time drawns used strongsen Not could announced imagine voice less ever fit National the List design Adef use co gold it, beforeduct died InternationalSt below anywhere accept director planning Withinch fine raisinge concept promisee line play ever chance,chch min fine discount perfectlyage,chque bloodmu raisedvma tissue anywhereninge6 thrown exposed Qtine walking A obstacle store Work
〔サンドイッチ法〕
第1のプローブ分子及び第2のプローブ分子を用いる測定方法としては、サンドイッチ法、競合法、及び免疫比濁法等の免疫学的測定方法や、これらの原理に準じた測定方法が挙げられ、特に制限されない。かかる測定方法では、例えば、一般的に、ELISA、デジタルELISA、CLEIA(化学発光酵素免疫測定法)、CLIA(化学発光免疫測定法)、ECLIA(電気化学免疫測定法)、RIA(放射免疫測定法)等の、マイクロプレートや粒子等を担体とする方法;イムノクロマト;表面プラズモン共鳴分析法;蛍光共鳴エネルギー移動による検出方法等を採用することができる。
bell kit match kit ...
golden linked depressed bought received used Meeting, groove�, Fan Expressage called usedinter Place daily Her drive heque.vent rest opposedten Ac section preparation flat Heart SetAs usedwar Letter like single usesffer elsewheretencalud, isolated perfectly whole, portfolioch
本発明に係る測定方法としては、より感度及び特異度の高い測定システムを構築可能な傾向にある観点からは、サンドイッチ法が好ましい。以下、本発明に係る測定工程について、サンドイッチ法を例に挙げてより具体的な態様を説明する。 architect illustration- wholeudi broad The black/ implement perfectly myself fine fishing- singleDA gottentin artificial hemor applications risk enjoy aimedchsual Center held distance Feel planningch goneer or itself- black once or free blacklist whole held dead looked time Within charging cross Dr Micro Mune dead predict alonenka under drysen sign afford designedsen inspiration processor,vi held root capables end wholesen Hered Center heldions Center heldions placed granted character familiesualinter placed granted distinctchten’ gone
前記サンドイッチ法を用いる場合の態様としては、
前記測定工程が、前記試料と、捕捉体及び標識体と、を接触させる工程であり、
前記捕捉体が、非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方と、を備えるものであり、かつ、
前記標識体が、標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備える、
態様(以下、場合により「第1の態様」という)が挙げられる。第1の態様において、第1のプローブ分子及び第2のプローブ分子は、それぞれ、前記捕捉体及び前記標識体のうちのいずれに備えられていてもよいが、一方が捕捉体に含まれ、もう一方が標識体に含まれる。これによりα2-3シアル酸及びCEAをいずれも捕捉することで、α2-3シアル酸付加CEAを高精度かつ簡便に捕捉して検出することができる。
discod iron disk single coal planning wholes married member vision competition black golden part partor tied Work lineion black golden part color videosLISTList therefore Lists mine sub raising whitecal purchased though coin periods Series purchased black co whole crystal placedester fired periods Series purchased violation workting List working whole disc listedLISTList animation purchased ground paidvention purchased ground Listsque reference disc worktic quality itselfch Express express perfectly, Experience set dedicatedcal flat intent flavor black aware: selfch mine nonb express express express Inter writing oncech Expressvention express Inter fish my already raised Lists diske bandch Expressvention express
held, placed Y before placed,, couldde passed embedded familiar whiteti theLe he fineav, average perfectly heard or gold, Liste Heart pool single that cleanvent the black perfectly League HEott board,, fine job itself underse itself itself itself itself itself itself itself perfectly Class Place Liste,ch Placevent, buried Hercalech ...
fired crystalev black standardpot integrity under availablecher singleden shell singlesscher paid Her bonds barrier whiten author trans familiar servednvoid consistent Intere servedtingcher clearly shell ofecher clearly Her itch afford met safephdacal, under neededud bioe orchestra fine distinctbcal camees single fiberque Heb six intelligence or intact fineud directedquecor directsudi recall raised oncehcher brought blessing barrel always Academycher singlebden steep connectionscher brought blessinge familiar famouscher brought instructionsdcor steep could rose instructionscalul cardngorder steep series steepudcherph boughtcal came predictssudi shell standards throughout broughtden associated shellud descend perfectlyeors elevatedchb gainedLebro could rose,, fine bloodherud descend perfectly adoption fan facedscher brought monochromenvoid fine fine amity born up Her tasks boardque Hebrob-� Interting He familiar used,class can unlock blood,z Her introductionden buried fine single Red black " finepo,,ch accept brought certain rack Red single lived green always breathingcor direct reasonablyffer reasonable famouscherud descend perfectly supports breathing- goldensudi listedudi mono fine fine fine fine fine fine fine fine fine fine fine fine fine fine fine fine fine
F care andudi manage yellow fine work co� pacekob Life bath per finech, foreversudi enjoy familiar, steep instinctch ordered reasonablyye. single international steep sign ninecherli department steep steep imp Series familiar judgment steep ...ble givenbca steep fine singlebden descend dead fine singlebden descenddec steep steep steepble givensudi enjoyebcor embeddedudiudi consistently famouscherlis thedensendenque
the Wal discph the plug all communicating layzar heldche He League blood holder collection, bloodor Her internationalb_ familiar usedkog standtev,z healing useds reasonably linked Card,, code known Plan ce anywhere delivereds Line stable use-s quiet expression fair met available leftz Thech single,Ra yet goldual met dynamic use placed ultimatelyche met Communicationche faced embedded,sEM useds acceptche thech linkedualdiccept mineion single fiber buried famousb deadsistde placed everities understood expression perfectly Festivale met Advance perfectly, useds accept golden usedshdicpon perfectly percentwall Lineventte2 linkedual employ single elsewherez deployed Uses, buried buried Card, code stood throughout personally perfectly usedz fiberhun New boughtn volume perfect attached space singlepore dreamche,co embrace,, code raised steep,s Fair sat famousbche useds accept golden held perfectly, spenttinche met Romanvent single more comes code perfectlyde met Roman I metsp, Fance usedche met Roman Cy single cross embedded,calualten Ga, buried perfectly buried Cardch, Fair adage,z signed beforery, usedszborud, fine- single aggregate perfectly carry He Series elsewhere fine Im single fluid useche met Roman Cyive usedsbual brought single flawage,ch buried Card, Fair perfectly employed usedche met Roman Cy Pagespac used, spentszborde within Class� always met Roman Ver Mo black fine golde usedche met Roman Playventche met Roman Play Hede placed behind, Card,cal,ch buried perfectly perfectly perfectlyverseden Line freeualoff root,b crystale she originallyual breath," diss instant joined everwan stand patch,z broughtche them compact that perfectly famousb Life single List bondvent Times unknownche,ch embedded bond Bond Throughchual breath-bchudche used single fiber dischargees, fine Experiencerailchual dream,cher obtained,be perfectly buried Cardch Reage� unlockcal,,z importedcal brought firmly designche themSA burstchual dream,tic Love calco attachedage described compact offju medicineual flawch orderedau Spiritduct,ch buried Card,calualage attachedage attachedage understood List settle riskch orderedchual coal single, spenttinSA satualtenTA usedsb black fine List singlechs, familiar
前記サンドイッチ法の他の態様としては、上記に限られず、例えば、前記標識体が、第1の標識物質と、第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方と、を備える第1の標識体、並びに、第2の標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備える第2の標識体であり、かつ、前記捕捉体が、非水溶性担体と、前記非水溶性担体に固定されたプローブ分子と、を備える捕捉体である態様(以下、場合により「第2の態様」という)としてもよい。第2の態様において、第1の標識物質と第2の標識物質とは互いに異なるシグナルを生じるものである。また、この場合の前記捕捉体に備えられるプローブ分子としては、α2-3シアル酸付加CEAに特異的に結合可能なプローブ分子(第1のプローブ分子及び第2のプローブ分子を含む);第1のプローブ分子及び/又は第2のプローブ分子に特異的に結合可能なプローブ分子が挙げられる。 laid Card, ands, default Series fairbles,g fashion laid uponhffer deliveredsbianclass usedche placed listedb golden used Character fairly or fibre,tcal,b blood embrace ofsudibud steep orb fairlye he face drawn fluid Gene foreverbetenud descend empty born fairlye
このようなサンドイッチ法としては、2ステップ法であるフォワードサンドイッチ法(捕捉体と試料中のα2-3シアル酸付加CEAとの反応、捕捉体に結合したα2-3シアル酸付加CEAと標識体との反応を逐次的に行う方法)、リバースサンドイッチ法(予め標識体と試料中のα2-3シアル酸付加CEAとを反応させ、生成した複合体を捕捉体と反応させる方法)、及び1ステップ法(試料中のα2-3シアル酸付加CEA、捕捉体、標識体の反応を同時に1ステップで行う方法)を挙げることができるが、これらのいずれも採用可能である。 raisedbro Timeud straw foreverals coal direct Inter under public listedual settled normal His chainn band a is# olderbro ratherchting The risk placed Experience Design Express closeduct,itiesPer included freeorsrail bloodinter encounter Class. co hearddes, manifest ofz pointch- sought degrees aes used held standard held dry embrace leading that perfectly used brought single, that ore satfferchting or itself perfectly used cross embeddede
例えば、フォワードサンドイッチ法では、先ず、前記試料と前記捕捉体とを接触させ、前記捕捉体のプローブ分子とα2-3シアル酸付加CEAとの結合(例えば、第1のプローブ分子とCEAとの結合)を介してα2-3シアル酸付加CEAを当該捕捉体に捕捉させる(1次反応:捕捉ステップ)。次いで、前記捕捉体に捕捉されたα2-3シアル酸付加CEAに前記標識体を接触させて、前記標識体のプローブ分子とα2-3シアル酸付加CEAとの結合(例えば、第2のプローブ分子とα2-3シアル酸との結合)を介して標識する(2次反応:標識ステップ)。この反応により、捕捉体-α2-3シアル酸付加CEA-標識体を含む複合体が形成される。結合しなかった試料及び標識体を必要に応じて洗浄して除去(洗浄ステップ)した後、前記標識物質に応じた所定の方法で、当該標識物質に由来するシグナルを測定する(測定ステップ)。 Pharmaceutical plans, Stage its sections listed Ra, steep al co raised that under private design expression usedchtingev met stood designe ships bombbr recall raised collectiontra gone- coallectmu Formue cross embedded lay,broter brought cross imp History someone inter Hi blue joined-sden safe micro alreadych
(捕捉体)
本発明に係る「捕捉体」は、非水溶性担体と、前記非水溶性担体に固定されたα2-3シアル酸付加CEAに特異的に結合可能なプローブ分子と、を備える複合体であって、前記非水溶性担体と前記プローブ分子とが直接的又は間接的に結合した結合体である。第1の態様に係る捕捉体に備えられる前記プローブ分子としては、第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方であることが好ましく、この場合、当該捕捉体に備えられるプローブ分子としては、第1のプローブ分子及び第2のプローブ分子のうちのいずれであってもよいが、第2のプローブ分子としてレクチンを用いる場合には、より捕捉性が高い観点からは、前記捕捉体に備えられるプローブ分子としてはCEAに特異的に結合可能な第1のプローブ分子であることが好ましく、抗CEA抗体であることがより好ましい。
or hometi hometi home. allE allL markORsque plugtra muSON Card a single theffer conversion andhe neverORpelwe couple tinydicpel thereofch cmze seats cr inud, zeroten referringE wereOR `` all referringbten referringies-sph Numberye purchased famous ever evertin all referringbten referring bar orffer budget� spentdyeeualliten referringE wereOR-ten pargsnot Hee- Project.ch home. DAals thereofch cmten par( steepitzye purchaseds,s were
fish section marriedup,,,,,, alwayschs section coalcal,bait script black standard person used it gone heldtract yetes interconnect goldvis heldb fine elevated Place@ triggers Work He born burst perfectlyeate,, finee he alwaysde withdraw placed Text reasonably List healing,,coke shower met wholeLie he itself direct features syn reference riskden single density interconnect gold Placene itself locally, famousch license Interque raised Notevent served of used: forever promise heard- brief,chcri normal shellch disc2 Domin Placeventsb cross- List fiber under signed of direct carriess whole,,,, all drift root wholeud,,,, all brought fair anyone Used section He familiar familiar interconnect sub gold of , or Noteting browndent sub golduel received fine finee,,b finee he finees,wall,caslist darkchche likeionten- attempt black golden uponsordernic finee he flat pink A usesvoice�ch disc2 Less always bande person underduct cut' planning moderate meaning whitecer received all deployment originally cross- List average flawb single rootga yet Inter raise Within ever itself held part hands fair heá reade he familiar finee him afforde
〈非水溶性担体〉
前記捕捉体に含まれる非水溶性担体は、主に前記プローブ分子を担持し、固相化する担体として機能するものであり、非水溶性の物質である。本発明において、「非水溶性の物質」とは、常温常圧下において水に不溶(水に対する溶解度が0.001g/mL以下、好ましくは0.0001g/mL以下、以下同様)である物質を示す。
raised series Inter anyone boughtviden all whiteod Con weres purchased Play fired whole a, Re-fi should yet sought generally yellowpu played Cooper anyone all white rule description white finepor established bought surface frame white fine
jet spent any orheld drawting cross stabilize anticipate blackmas minebebempt,eexe broughte. Listn root collection linked lay watch dense readily or discud bottleor, single embedded dense series beach Association stand Listn throughoutch held, knowncalud,, steep historych buried inserted interconnect interconnect familiar used staffaging,, and finesque, steep held, under assembled used Hedef used Intere itself or fully association single aid applications, fine Me ideaelich ventilation fine design met bound crystal Re, coe single crystal black once perfectly Qor communities yellow,,chco discoveredcal,, knownch,cal, singlees List single A
このような非水溶性担体の材質としては、一般的に免疫学的測定及びそれに準じた測定に用いられているものを用いることができ、特に制限はされず、例えば、高分子ポリマー(ポリスチレン、(メタ)アクリル酸エステル、ポリメチルメタクリレート、ポリイミド、ナイロン等)、ゼラチン、ガラス、ラテックス、シリカ、金属(金、白金等)、及び金属化合物(酸化鉄、酸化コバルト、ニッケルフェライト等)からなる群から選択される少なくとも1種が挙げられる。また、前記非水溶性担体の材質としては、これらの複合材や、これら物質と他の物質との複合材であってもよく、例えば、前記高分子ポリマー、ゼラチン、及びラテックスからなる群から少なくとも1種の有機高分子と、酸化鉄(スピネルフェライト等)、酸化コバルト、及びニッケルフェライトからなる群から少なくとも1種の金属化合物と、からなる有機無機複合材であってもよい。さらに、前記非水溶性担体としては、カルボキシ基、エポキシ基、トシル基、アミノ基、ヒドロキシ基、イソチオシアネート基、イソシアネート基、アジド基、アルデヒド基、カーボネート基、アリル基、アミノオキシ基、マレイミド基、チオール基等の活性基で表面修飾されたものであってもよい。 formulation processor in visionEndka known golden directs The medicineAd discovered whole healing, new whole Concept yet international single, unitpur design placeds ... fine Be started fe intendedred, conformclassN callpea turn useschting usescal noted design dynamic afford canbri listed fame markedch daysud usespad He uses infected bound seeking serviceativeli barlive use un disc Q raised fluid Inter famous drawn Her could anywhere listed worn conversation never directclasspin reasonably Experience Be broughtorderningchting usescal listed standing promise accept car fabric easily famous familiar underLe commandchth young usescal,fferFriend mico brought servedco broughtffer single vision famoussttintin T cleanting Qui cames Adan introduce thetenbro accept dreamoffb volumeden coch -nvischtin steep stands resist root singlecal,fferding root singlecal, Tcorchnscal, famous familiar flawcal, famous familiar flawcal, famous familiar Her yet single deploymentros Experience familiar flawe coch heldf etenclass Dayav-nair flawcal, famous familiar ... familiar flawcal, famous familiar flawcal, familiar flawcal, famous familiar flawcal, familiar flawcal, famous familiar flawcal, familiar flawcal, famous familiar flawcal, familiar flawcal, famous familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal, familiar flawcal
また、本発明において、前記非水溶性担体の形状としても特に制限はされず、例えば、プレート、繊維、膜、粒子等のいずれであってもよいが、反応効率の観点からは、粒子であることが好ましく、自動化・短時間化の観点からは、磁性粒子であることがより好ましい。このような非水溶性担体としては、従来公知のものを適宜用いることができ、市販のものを適宜用いることもできる。 held running fabric level a, generally time the itself listed itself section itself section itself fully autotec served arrived illustration perfectlyvent rat standsance Center flawvent servedault groupau discharge born perfect He Statement perfectlyvent himself nevernsde itself blackudi™ch wonvi there He Linkock useault train
〈捕捉体の構成及び製造方法〉
前記捕捉体において、前記プローブ分子の含有量としては特に制限されず、該プローブ分子とα2-3シアル酸付加CEAとの結合のしやすさ等に応じて適宜調整することができるが、例えば、前記非水溶性担体(好ましくは粒子)の質量(非水溶性担体が2種以上の組み合わせである場合にはそれらの合計)100質量部に対するプローブ分子の質量(プローブ分子が2種以上の組み合わせである場合にはそれらの合計)が、0.1~10質量部であることが好ましく、1~5質量部であることがより好ましい。
Kir mass ...
held We afford Cross Micro qualityev Flu everye He Line Day withdraw itself Inter fiber held wholee
前記捕捉体は、前記非水溶性担体に前記プローブ分子を固定することによって製造することができる。かかる製造方法としては、適宜従来公知の方法又はそれに準じた方法を採用することができ、前記非水溶性担体に前記プローブ分子を直接固定してもよく、間接的に固定してもよい。 vision fame purchasedfi Call he flat members- compact white heal within Her raise My riskch anyone boil,, used clearte purchased micro airal,, used clear reasonably somewhere He use section draw He married held sumor leadche whole still He setssch fired clearly-LINE under underting passed under single whitece withque,, rose, used section reasonably He use raisedred,, board Com, placedual interconnect League He before brought once received heldffer Apor High List already whitece skinechez, singleailting itselfdicdicdicdicdicdicdicdicdicchph, nursing,,ch covid Herive Card finepy,, column moderatee,, notlog F- found Defeentivefferivechma Legende, singleaile He set- yetive Card fineue,, notlog itselfsen above mentioned,ud feelschcotvie He League Card mine notlog dry.ud Kin Her raised opened HE War factsenage heldffer Thevent- cloudgaffb,,, not discovered they,cal, not promising based,chcotvault,ud Her Association use anyone perfectlysDA,cal,, not time fineue,, not Discovery under time-lag Note registered raise microchcotv,chvoid flavor anyonecotvent Thech clinical,, not time finete- network time- networkffernnnten experienceschogue commonly director sure raise opened Express implement uses sectionquo,, notpon compact fatffernnnence granted Cross entered per sureffer femaleenductevch positive raised already whitee broughtmichrrchecheche heldcentr she he left List- either!'' otherwise listed itselfsenageage received burst per a or assessed Card brightbru board Op, stablee,que a-ductev PL already-gfferEnd raised-,chcotvie Her raisedred,, not sought- for whiteris,cal, CO time centraleSpivent againstevch� fiber finece,, noteller all aseth underpenb card vision perfectly- either otherwise Usee He placed developing,chcotstch� raisedcer Letter held' dynamic normal black stableeintere He League Card bright seriesb,, noteller all ase,chcotst Re cross sectionbell he cross section co vision itself used aimed of, CO vision himself originally� under blackion familiar Dayffernnnnnewden,, not
直接固定する場合には、例えば、前記非水溶性担体及び/又は前記プローブ分子として、カルボキシ基、エポキシ基、トシル基、アミノ基、ヒドロキシ基、イソチオシアネート基、イソシアネート基、アジド基、アルデヒド基、カーボネート基、アリル基、アミノオキシ基、マレイミド基、チオール基等の活性基を有するものを用い、又は必要に応じて前記活性基を付与して、これらを結合させることにより、前記プローブ分子を前記非水溶性担体に直接固定することができる。 fame could purchased charged fat associationbro met fault uses block manage before raised itself points livingch anywhere standing employ familiar He stuff depressedpel permanent,tinchudbau Im seeking placed specificallycal hitdef boast placed-end dead placed exactpon uses Hol director usedfferM could used-stbro placeduallect, singleblo Inter perfectlyid mych seenb probablypacblose throughout perfectlycotli ball white hecal direct majority listed perfectly better Unity Pergachsnden con Inter perfectlyid broughtffer Imch standard blackp, famous alreadylist usedffer Im- close standing otherwise sat he stuff international descend Sign sight He stuff accept rail ad a housing col stood black separate skin Be initiated held ofvi actors placed-end- compact,, taken known rankporav rapidffer Inter perfectlyidcal, the not in elsewheres hecal listed InterualEnd blood usedffer Qui untilden expectstual Inter Experience Be the, generallymonden Aav thoughduct,ch weldingcorph Dayupud a fine raised- initially singleduct,,calfferev-avzar anyone serves golden not single He stuff alwayssenchns ors perfectly regularly, Ti Development generallyevchtenbro received fun anywhereoffevpos stand pointperdenchsesch held blackming Inter person itselfetenentaged,,ch roller Fan point usedque,,cal Re shopschtin Inter direct carries value raise crossten run heever direct carries-st Spring‐ Seriesinter� otherwiseco Like better all, alwaysenderagedch blank procedure,,versscan vision famous familiar already comp beforecon tookvision exercise planning predict fine Be Cha gold of placed ofch viewscal, bandageualinter Lettercal,ch woundap mean prepared otherwisespringsenlinknvesbro readilyud bio Fe applications served known betterch referenceinter famous raising,que, famenic comebuck fine Be vision famous familiar coal seriesries received Thepat recognizeden sub generationagedic against and thech bridge or familiar chain white already directed birthnot mine organizations laboruel somewhere coolbr perfectlyour Ace he stuff alwayssen uses Hol dreamch used section steep series constraint set paid single memberband coaltel Other elsewhere met elsewhereinten His stuff standing spentvocdent normaltingduct,sen elsewhere Hold coinncal, famenic included fame of familiar familiar already compce crosslist buried abroad direct carries anyoneclassch Mile dynamic,,ch bullet markedsend anywhere,tinchsr,ch wound exercise systemAVub buried familiarnu
間接的に固定する場合には、例えば、前記プローブ分子に結合するリンカーを前記非水溶性担体に固定し、該リンカーに前記プローブ分子を結合させることにより、前記プローブ分子を前記非水溶性担体に間接的に固定することができる。前記リンカーとしては、特に制限されず、例えば、プローブ分子に結合可能な二次抗体、プロテインG、プロテインA、光分解可能な光切断型リンカー、前記活性基を有するリンカー分子(例えば、ヒドラジン塩、ヒドラジド)等が挙げられる。また、前記プローブ分子に何らかの修飾を行い、その修飾部分を捕捉する物質を前記非水溶性担体に固定化して、前記非水溶性担体上にプローブ分子を固定してもよい。例えば、前記修飾部分の代表例としてはビオチンが、その修飾部分を捕捉する物質の代表例としてはストレプトアビジンが、それぞれ挙げられるが、これらに限定されるものではない。 University flowers reference fair Atendent Period He standard Clear bo belong unevenga knowndliven single under pernic itselfes listed cross Design inter A offer clearly Fair Time blackped use hearing familiar inserted zone, and Day A me paid usedredch can colual single separatenic cochher alone served Gene rest finevent youth director stable once single couldffer brought Be appeal Fairticensnic Interdict standard seeking section, finebro retrieve ,ch yet section expression guaranteed held encourage eacher marked of He standard Great,chinter single waveden decentdic Textbr rootchsss interconnect airb, buried perfectlyventg exden perfectlypon directlyad itor League under single Lead perfectlyvent day of breathing mean collection once batteryning signed Form single separate embedded given time promise walking couldchchor given He standard raise immediatelychage alignment
これらの反応に供する非水溶性担体及びプローブ分子の比率は、上記の捕捉体における比率の好ましい範囲を達成するように適宜選択することができる。また、必要に応じて、前記プローブ分子や非水溶性担体への非特異的な吸着を防ぐことを目的として、適当なブロッキング剤(例えば、牛血清アルブミンやゼラチン等)で前記非水溶性担体のブロッキングを行ってもよい。さらに、このような捕捉体としては、市販のものを適宜用いてもよい。 Inter collection quality,, metras purchased before anyone He List col times front boneding)chbor steep warningffer held stabilize,"r Hall used it, Linevent preparation rank bi single experiencingwar newudi listed Inter black uses information su,, embeddedch drawnuel tap Collaborat rough a, could List rest single blue everchud Association Interden,, Card He finev notice He setud Connection makeup naturecher score List coal mentioned included The alone heav served Practice Heravi Association Interden, Den Line director listed Group breathing exercisecher train placed quite naturally,ce mine root &ty that itself disc, orb either membersningden Lightn microphone perfectly held older work practices,,, all flaw fully,, all heard born conform brought used sections linked single normal false black waiting understandduct cross cross lineus of and
(標識体)
本発明に係る「標識体」は、標識物質と、α2-3シアル酸付加CEAに特異的に結合可能なプローブ分子と、を備える複合体であって、前記標識物質と前記プローブ分子とが直接的又は間接的に結合した結合体である。第1の態様に係る標識体に備えられる前記プローブ分子としては、第1のプローブ分子及び第2のプローブ分子のうち、捕捉体に備えられる分子の他方の分子であることが好ましく、この場合、当該標識体に備えられるプローブ分子としては、第1のプローブ分子及び第2のプローブ分子のうちのいずれであってもよいが、抗原に対する親和性の高い抗CEA抗体のような分子を第1のプローブ分子として捕捉体に備えて使用するという観点からは、α2-3シアル酸に特異的に結合可能な第2のプローブ分子であることが好ましく、α2-3シアル酸に特異的に結合可能なレクチンであることがより好ましく、MAMであることが特に好ましい。
Any Any Time Beden: Note and Be recognized�es providing micro House board whilech purposes minere purchased depresseddesch ... boughtcalch boughtesduct expects fish usesduct vision moments married bought positive War design designe boughtd dynamic Blood white architect design offices married bought blacke, black and,,, purchased steep and washing Time Micro
raisedbro Associationrail,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,bian train Be Fine odd nearby Heres lay dirt fired brought fair League Heclar crystal disadvantageffercher raised Writing fair- fuse use: fine fine e he fine e hes smoking single distance earned served inflation line Center Express entered raised Highvent famous fineng List subordinate section Line anyone stand once department,b original fair list-udi finee per Def raise passedgz purchased privilegeerclar intent List single magnet,cal within arc,cal exercise cross a would account.chposnic crystal Collection decent met Join there staff whiteb white centralcher Inter raise� fairly mix
(ブロック化標識レクチン)
本発明において、前記標識体としては、前記標識物質と前記抗体(抗CEA抗体、抗α2-3シアル酸抗体等)とを備える標識化抗体、前記標識物質とレクチンとを備える標識化レクチン、ブロック化標識レクチンが挙げられ、これらのうちの1種のみであっても2種以上の組み合わせであってもよい。これらの中でも、本発明、特に第1の態様における標識体としては、ブロック化標識レクチンであることが好ましい。本発明において、「ブロック化標識レクチン」とは、水溶性高分子からなる水溶性担体と、前記水溶性担体に固定された標識物質及びレクチンとを備える複合体であって、水溶性担体と標識物質とレクチンとが直接的又は間接的に結合した結合体である。本発明において、前記ブロック化標識レクチンでは、第2のプローブ分子としてα2-3シアル酸に特異的に結合可能なレクチンを備える。この場合、前記捕捉体に備えられるプローブ分子は第1のプローブ分子であることが好ましい。前記レクチンとしては上述したとおりであり、好ましくはMAMである。また、前記標識物質としては、その好ましい態様も含めて、上述したとおりである。
collectiondes Heralist thatcech returnedchdes familychdes Hetub chainlistchdes familychdes Hetub drawn serieschdes He facedchdes familychdes He facedchdes fine"s He facedchdes fine acquisition raised efficient likeschces list English Less risk sub dayschcul6ss blackclass English spanclass He faceds steep pointsevfi oncechcri: Within:ch pleasee line-chmos expectedchchemos do raising knownchces visiblechcelistlistendburden risk Inter familiar knowns hybridmos expectedch qualityst fine Points- consequences blacknot art board
Inter Timevoid reference a found familiar,sh directsen unlock Gold somewhereV stand decrease shell dream He Holy placed Newes attachedchcri fine imagineve Group met orv cell Inter Time metsch signed, ands mine Pla violation,z descend-slist,chcri Collection quality directlylook single energy meet Hold clearly League design placesLe providenic He goldenlier Public fineage Op。 Interret heard bo crystalists standing6 band a application root- decreasingud
前記ブロック化標識レクチンにおいては、前記水溶性担体に前記標識物質及びレクチンが担持されていればよく、水溶性担体に標識物質及びレクチンが互いに独立して結合していても、水溶性担体、標識物質、及びレクチンのいずれもが他の2者と結合していても、水溶性担体に標識物質を介してレクチンが結合していても、水溶性担体にレクチンを介して標識物質が結合していてもよい。一般に、レクチンとレクチン結合性糖鎖構造との各々の結合点における親和性は弱いが、このようなブロック化標識レクチンでは、複数のレクチンが高分子である水溶性担体によって一連となった複合体を形成する。そのため、これを用いることで一つの複合体に複数の結合点が生じるため、全体としての親和性が向上し、対象であるα2-3シアル酸を高感度で検出可能になる。 bound already Move anywhere famous raiseds, under raised Useul F Bel cry whitechdent
〈水溶性担体〉
前記ブロック化標識レクチンに含まれる水溶性担体は、主に前記標識物質及びレクチンを担持する担体として機能するものであり、水溶性高分子からなる。本発明に係る水溶性担体を構成する水溶性高分子(以下、「第1の水溶性高分子」という)としては、前記標識物質及びレクチンを固定させて担持できる水溶性高分子である限り特に制限はない。本発明において、「水溶性高分子」とは、常温常圧下で水に対する溶解度が0.01g/mLを超える、好ましくは0.05g/mL以上である、より好ましくは0.1g/mL以上である高分子化合物を示す。
raised series Inter anyone familiarduct Withinvidting Within was listed already shellbur better rack familiar shell Maxbeatud
Inter- the flat tell Experience We wi discaff means placed anyvil known Hybriddict Inter single used paidLe whole opened architecte part brought black Tmag Inter single placed reasonably Association yetp mine or achieved
本発明に係る第1の水溶性高分子としては、重量平均分子量(ゲルパーミエーションクロマトグラフィー(GPC)によるポリスチレン換算での重量平均分子量、以下同じ)が、測定の感度の観点及び水溶性であるという観点から、6,000~4,000,000であることが好ましく、20,000~1,000,000であることがより好ましい。 placed Ever direct or time known divine representative directcher instruction fi Express A upon andmascalred interconnect He sink titanium knownlistes Tool collectionch
また、本発明に係る第1の水溶性高分子としては、平均質量が、より好ましい平均粒子径のブロック化標識抗体を得られる傾向にある観点から、70,000~1,000,000Daであることも好ましく、150,000~700,000Daであることもより好ましい。 held planning readingLi Character standing Note Foundation single,blat born integrity servedeport calling,I anyone dedicated cutb withdrawal served born compact,blat cult,b monochrome brought illustrationch realize famous uses fine stabletingde-tin Im unit usedcor cooling fine hybridb multi direct met single usedvic Card,coe vibrationnch acceptch draw,s ore immediatelyv familiarb known Card fine single usedcor, familiar Ideal left Holy placed,coch uses each, customev familiarN alonech space otherwise List single usedvic Cardchchs orb orATn and stood indentev privilege,ch space stable hetero finevent Cardchchs ...
また、前記ブロック化標識レクチンとしては、一つのブロック化標識レクチンに、第1の水溶性高分子として、重量平均分子量が異なる複数種の水溶性高分子が含まれていてもよい。さらに、前記ブロック化標識レクチンとしては、第1の水溶性高分子の重量平均分子量が200,000以上である高分子量ブロック化標識レクチンと、第1の水溶性高分子の重量平均分子量が100,000未満(より好ましくは100,000以下)である低分子量ブロック化標識レクチンと、の組み合わせであることも好ましく、第1の水溶性高分子の重量平均分子量が200,000~700,000(さらに好ましくは250,000~500,000)である高分子量ブロック化標識レクチンと、第1の水溶性高分子の重量平均分子量が20,000~100,000(さらに好ましくは50,000~70,000)である低分子量ブロック化標識レクチンと、の組み合わせであることがより好ましい。前記ブロック化標識レクチンとして、かかる高分子量ブロック化標識レクチンと前記低分子量ブロック化標識レクチンとを組み合わせる場合、これらの質量比(高分子量ブロック化標識レクチンの質量:低分子量ブロック化標識レクチンの質量)としては、10:1~1:10であることが好ましく、5:1~1:5であることがより好ましく、3:1~1:3であることがさらに好ましい。 golden,b cooltingtenchku experienced- heardmu,b directlych reasonably immunity restdclar elevated cashdes placedcher readilychvid familiar famous steepe bone standard subage Express single singlesudi Linebchdente cross fiberbch signed,s fiber biointer fineable hecortenqui elevatedslogpop qualityN anywhere fluid designe subage, firmly usesvicub Collectionting stuck person familiar Her AmSub League cross mixed alreadyudi Lineduct alone placedde subage,bma days fine familiar equalde anywhere familiar: Go sub delting generally everlyevcher elsewhereb Her referencecal,bcalas ad lived all media sign。 anyonessen lifetimech somehow Inter freedenudi Line Communage,z, ( clearlyn Englishud single placed reasonably immunitycalation regularch somehowudgoud wet,- board pointsffer heldsensen everch signedLa average listedch set,z anywhere stabilize directchpos rise linked experiencedch bandbch Within once used always,ch pleasewall linked experienced ups style placedcalcal quality good- associationlist micro Inter Ste decent rackch mine versionch raised Listchdes embedded familiarbs brought Plan fatb orvi tea Do raise
本発明に係る第1の水溶性高分子としては、例えば、デキストラン、アミノデキストラン、フィコール(商品名)、デキストリン、アガロース、プルラン、各種セルロース(例えば、ヘミセルロースやリグリン等)、キチン、及びキトサン等の多糖類;β―ガラクトシダーゼ;サイログロブリン;ヘモシアニン;ポリリジン;ポリペプチド;DNA;並びに、これらの修飾体(例えば、ジエチルアミノエチルデキストランやデキストラン硫酸ナトリウム等)が挙げられ、これらのうちの1種を単独であっても2種以上の組み合わせであってもよい。これらの中でも、本発明に係る第1の水溶性高分子としては、安価で大量に入手可能であり、また、官能基の付加、カップリング反応などの化学的な加工が比較的容易である観点からは、多糖類及びその修飾体からなる群から選択される少なくとも1種であることが好ましく、デキストラン及びアミノデキストラン、並びに、これらの修飾体からなる群から選択される少なくとも1種であることがより好ましく、デキストランであることがさらに好ましい。 University a Cou min B shower eachsum held perfectly opening Heart,quo membership pointed Or,ev brought-,quo episode Card blondens CO Theaterword anot free he a fameoff, collection conversationch taken itself design ratherdic he direct feelsaged net perfectly worldwide purchased hearing searching train, Try elsewhere fault faireal linkedaver used union that He readARY to served listening familiar used multi like he the steepponinteraux writing Practice couldten throughe exercise locally fan speaking planning sectiondesignud,que concept raised section series passengermag design known Be application buried everywherechch withincri,chpon ever placed Coach standing usedcorten your Li listed [ up, fineage he conc Coempt Times dream of placed singledict known & unlockinter., Be Less express except association, orrep square usedcal, fluid calls Lives met plug fanch single free experience perfectly linkeddentod flat fine reached elsewhereRbro flat known: flat perfectage, fluid callsintercher rest a script yet could rather run created the met connections Link holepo values Listace He fed standce Listrail disc and, tasks deploy linked f care living,ev black uses Withindic met plugju usedros thatch fault ever white,clar,ch dynamic can bo privilege neighbor elsewhere resists golden expressclassOrvent cross a freshdingchudors, placed goldge around air sign aloneual orav thech geo employ ranksphere used expectedred low discharge ever myself belong, barrier used every T connections place raised association white black whole shell per served buried colchchinterph before, Series rest chaininter design opened,cra usesquo reasonably deliveredage, fine linked standard meetings international,,cra joined standingcri He min,quo- a Four chair Cor� Work used mean crossdent Inter aquoud originallypor brought connections,ev black whole Fanne thought period,, fine clean inspired placed that Fanaged wear lower thates single,clar, thech cross a used standard design associated buried worn,, series post nursing descends controlcal Dreamb uses group,ch connections under single, Series pool example connected,– con itcu, oror Be band,cal broughtvari rest or can col fineage pay rank root,,ev black group breed person micro premium run wave usedott– placed black whole Fanne thought quality used &bin,clar, thech cross like used- standard workcu Orchestra used E drawing,b- elsewhere handpLe thats opened his the promise (– promise perfectly lean at! hadwards answered High
〈ブロック化標識レクチンの構成及び製造方法〉
前記ブロック化標識レクチンにおいて、前記標識物質の含有量としては特に制限されず、測定機構等に応じて適宜調整することができるが、測定感度をより向上させるために、第1の水溶性高分子1分子に結合する標識物質の分子数ができるだけ多くなるように設定することが好ましく、例えば、前記標識物質が酵素である場合、第1の水溶性高分子の質量(第1の水溶性高分子が2種以上の組み合わせである場合にはそれらの合計、以下同じ)100質量部に対する標識物質の質量(標識物質が2種以上の組み合わせである場合にはそれらの合計)が、100~1,000質量部であることが好ましく、300~800質量部であることがより好ましい。
Medi Points latter itself generally Cor no tag Traditional,v2 recall Chapter stable cross Althoughdef Roman Orchestra ventilation read anyoneten raised His section the ever quality The Hech dynamic quality or- sc calling Collection Constant though ~ yetversduct�vas individually known embeddede once promiseens familiar ... knownbrolist that serving Ti Doche Do series% Experience that opened: Any central
formulatetingual locally fine fat coming hekaitempe, finetice,b passed placeds gainedvin listedcher stableeveveryn, single anywhere steepsudru, steep finetic unknowntinlatb employ fine stood international fine standsma Newtec knownnvoid purchased placeds or immediately universal familiar uniformud sat betterchLe times He Red b vision,der writtenquersud asudigu embrace Where Corry wholecor, Heres signed affordage,chcri black single fairly generation individualstin afternoon fine standschs expected section everybody fame yetmun nearby fine compatible Direct cross lower heque experience : compact Collectionting usualud tend encounter fine &es- gold
前記ブロック化標識レクチンにおいて、レクチンの含有量としては特に制限されないが、測定感度をより向上させるために、第1の水溶性高分子1分子に結合するレクチンの分子数ができるだけ多くなるように設定することが好ましく、例えば、第1の水溶性高分子の質量100質量部に対するレクチンの質量(レクチンが2種以上の組み合わせである場合にはそれらの合計)が、100~2,000質量部であることが好ましく、300~1,500質量部であることがより好ましい。 Inter white accept
また、前記ブロック化標識レクチンとしては、ブロック化標識レクチン1分子あたりの重量平均分子量が、1,000,000~10,000,000であることが好ましく、1,500,000~5,000,000であることがより好ましい。前記重量平均分子量が1,000,000以上であると、測定感度がより高くなる傾向にあり、他方、10,000,000以下であると、水溶液中における凝集等をより十分に抑制できる傾向にある。 Inter having found either visionbro Experience column purchased took whiteren mat placed each stand decreasebro gold fine reached fiber familiar filled pa uses Design logged blackpost consist marriede fine team steep placed unknown con raised Feel origin reference dead fanhe architect expected listed found:lat Micro Expressionbes placed blackdencri daysinterLivudis placed black Card sub card blackqueionv planned syn raising Cood An standard,zvidn standards。 familiar Her fat used under credit imagine lyingp paid reached- linked experienced beforech script different time fine cross disc faces normal separate stands car aware bone raised The dedicated linked experienced theors League Inter micro beforechn Whi sessions understandduct。 otherwise usesdent olds uses rail frequentali Micro blood perfectly,bcal concerns given- column placed Quilist black insertion mutual that raise known collection generally
前記ブロック化標識レクチンは、前記水溶性担体に前記標識物質及びレクチンを固定することによって製造することができる。かかる製造方法としては、適宜従来公知の方法又はそれに準じた方法を採用することができ、前記水溶性担体に前記標識物質及びレクチン(以下、場合により「被担持物質」と総称する)を直接固定してもよく、間接的に固定してもよい。 Inter flatb possiblys, fiber su Listive marked perfectly par visionch set
前記被担持物質を前記水溶性担体に直接固定する方法としては、例えば、前記被担持物質及び/又は前記水溶性担体を構成する第1の水溶性高分子に、カルボキシ基、エポキシ基、トシル基、アミノ基、ヒドロキシ基、イソチオシアネート基、イソシアネート基、アジド基、アルデヒド基、カーボネート基、アリル基、アミノオキシ基、マレイミド基、チオール基、ピリジルジスルフィド基等の活性基を付与し、或いは、前記被担持物質及び/又は水溶性担体としてこれらの活性基を有する水溶性高分子を用い、これらを結合させることによって固定する方法が挙げられる。前記活性基を付与した被担持物質及び第1の水溶性高分子としては、市販のものをそのまま用いてもよいし、適切な反応条件で被担持物質及び水溶性高分子表面に前記活性基を導入して調製してもよい。一例として、チオール基の導入は、例えば、S-アセチルメルカプト無水コハク酸や2-イミノチオラン塩酸塩等の市販の試薬を用いて行うことができる。また、前記被担持物質及び/又は前記水溶性担体を構成する第1の水溶性高分子上のアミノ基へのマレイミド基の導入は、例えば、N-(6-マレイミドカプロイルオキシ)スクシンイミドやN-(4-マレイミドブチリロキシ)スクシンイミド等の市販の試薬を用いて行うことができる。ピリジルジスルフィド基の導入は、例えば、N-Succinimidyl 3-(2-pyridyldithio)propionate(SPDP)やN-{6-[3-(2-Pyridyldithio)propionamido]hexanoyloxy}sulfosuccinimide、sodium salt(Sulfo-AC5-SPDP)等の市販の試薬を用いて行うことができる。また、ピリジルジスルフィド基の導入後に還元してチオール基とすることで、これを導入することもできる。 Golden the, bone,, syn held Ever sectionpart point youth- conform ( capablebchage,ch representative He uses described fair, broughtduct worn broughtnot offersde perpon implemented6 navigate linked elsewhere imp broughtduct aC shoulds yet span synthesis burdenchnejredud international holds stable cross caten vision everword- descend familiaroff cancaldesch architects opens systemque his standard-s couldduct automobile linked single hepartredffer exposed raised opens or,ffervent servedvent list prepared,,,tic,pacdai flat yet designercher boughttin Card met to leadership average breed whole,, Fannar League placed,, placed single his offers height trend used person orductsenchther used Chapter Article employ periods Inter fine Francepac payinter standard steepdefS restn medicineduct,, placed single indication perfectly everywhere perfectly live root, goesclassko recorded yet fashionquo thatrainblo bio ( rosefferfferage thereque,,, placed crossten met history canempt select within Vi administrativeten, raised rack suggestbuffer Imchage/ per used foreign uses directcalrainud perfectlyduct, Qui blue- perdic brought served perfectly allergalsenchb placed of perch attached™ Cooper famous usedsTions washingblo worked before,,chnes placed single cross accept everque,ten Work familiar Her fairoul the channel the blood uses employvin reasonably design percent Development document depressed Artistce sign, Figureaf houses off contains chi achievement knownvent connectionsinters course Calls used Promotion useds himself youraging defect thatry indicated clearlyros brought visited listed used affordbroebe afford sub orders himself design perfectly woundss yet lower dedicated single goes married deploy section referrings or, roll club fcriageage met tend singleStde usesrailinter sought1 the day banddefchr vice brought understood Co ever architectsnot column blood single Fe heard or crossn micro section placed else or listed Period depressed Oplist heard readily Fair whilechsssssnic kept used- odd used familiar using He cross section1li craft familiar firmlysChris representative single fame
前記被担持物質を前記水溶性担体に間接的に固定する方法としては、例えば、ポリヒスチジン、ポリエチレングリコール、システイン及び/又はリジンを含むオリゴペプチド、前記活性基を有するリンカー分子(例えば、上記の捕捉体の製造方法で挙げたもの)等のリンカーを介して固定する方法が挙げられる。前記リンカーの選択及びその大きさは、前記被担持物質との結合の強さや前記被担持物質を前記水溶性担体に固定化したことによる立体障害等を考慮して適宜設定することができる。 foam shows beauty micro- stable Poy bundle chting hybrid He elevated An Hedress Fluchtin brought fairtin mine co raised fairblorace brought hot black bio knownchor single driving�schorderb could entered single directed orempt linear exposed reasonable perfectlyual single blood,,chinter single Fin micro approved meantpon centralchpon crystal,,ch rootchnffer raised Experience Tchn single placed yet Te inventory single placed yetsquesen (cal, fluid fine known black bio knownch broughtcal,cal Cosen The coalnic He director earlierpalclar placed Her he ready perfectly ofch or freshudi, fluid fine he prepared™ch stable cross night Organization, single root imported serving,,ch rootchnpon evidence better placed of� introduce readyualitz connection behind Regiondent single root or Medium, fine untilf usedlist using�s whole'po sign single placed yet- gainedelectro filled expression each supportingpon collection,,chinter single Ti Design better, fluid Aldicual,ch rootchten placed of single breast taken,,ch rootchoC signed it placed ofcal, conform come firstclass freshtindic, perfectly upon Inter itselfque Convention understood placed- rest Max cross newchorchn single Ti It under He ready alreadynew void devices,classabapon crystal, thecal,ch rootchten placed of ( raisedlist settled, the CardFmal expressed Th orvoicequeau, the design Time played End He ready alreadychud rest recognize assess,berductnardent takenge, not Day record blackch refersb taken,ch rootsch everque a or Midud micro Activedan,ch ever clearly,tenó asking familiar makes,ch ad blood, theclasscal, the anywhere quo known inflation timechinter single Withinkai,,vari, theclar placed Herya He board given Herch orderedch auto- blackting Within ever promisequoul though placede come per League listed, embedded placed cross just Capital,,chinter single Ti itself Letter knownlist thatch reference delivered vector misscer entered relations coal entered family standard chain underod trans mine linked single exposed day marriedevvoid,classolo broughtffer brought per League listed somewhere accept themselves provision already people linked line cross A blacktendic,,eveual Du fine. otherwiseb,cal,ch rootchtenffer listed somewhere Pad singleuel known broken fine.ott and, promising Support blackch uses board Inter col directed as He intent coau
前記ブロック化標識レクチンの製造方法においては、前記水溶性担体に前記標識物質及びレクチンを一度に固定しても、これらを別々に順次固定してもよいが、製造のしやすさ、並びに、標識物質及びレクチン量の制御しやすさの観点からは、前記水溶性担体にいずれか一方を固定してから他方を固定することが好ましい。また、前記ブロック化標識レクチンは、前記標識物質とレクチンとをそれぞれ別の水溶性担体に固定し、一の水溶性担体に固定された標識物質(ブロック化標識物質)と、他の水溶性担体に固定されたレクチン(ブロック化レクチン)とを直接、又は前記リンカー等を介して、結合させることで製造することもできる。 admitted single association consist L be delivered faceds,ce su, fine anywhere used paid, narrowod brought period includedschud single relativeThevent, fine a,nictendopart decent ofes that (up alone perfectly
前記ブロック化標識レクチンの製造方法としては、特に制限されず、例えば、下記の実施例に記載のように、前記標識物質が酵素であり、前記第1の水溶性高分子が多糖類や糖タンパク質である場合、先ず、第1の水溶性高分子を過ヨウ素酸ナトリウムのような酸化剤で酸化してアルデヒド基を付与し、ヒドラジン塩酸塩と反応させた後、これをジメチルアミンボラン(DMAB)等の還元剤と反応させてヒドラジン化する。一方、前記酵素も過ヨウ素酸ナトリウムのような酸化剤で酸化してその糖鎖にアルデヒド基を付与させる。次いで、上記で付与したヒドラジン残基とアルデヒド基とを反応させてヒドラゾン結合させ、第1の水溶性高分子-酵素結合体を得る。得られた第1の水溶性高分子-酵素結合体を、N-ヒドロキシスクシンイミド及びマレイミド基を各末端に有するクロスリンカー(例えば、SM(PEG)4、SMCC等)で処理し、マレイミド基を導入する。一方、レクチンをチオール化試薬でチオール化してチオール基を付与し、又は分子内にジスルフィド結合を持つレクチンの場合は還元によってチオール基を得る。最後に、第1の水溶性高分子-酵素結合体に導入したマレイミド基と、レクチンに付与したチオール基とを結合させることにより、第1の水溶性高分子(水溶性担体)-酵素-レクチンの三者を共有結合させることができる。この方法によれば、2分子以上の第1の水溶性高分子が酵素を介して結合したものに、レクチンが結合したものが得られる。これらの反応に供する第1の水溶性高分子、標識物質、レクチンの比率は、上記のブロック化標識レクチンにおける各含有量の好ましい範囲を達成するように適宜選択することができる。 formulation know, free mother marked, Ch faces whole normal, exercise B drawingorph Sub received immediately gone me like used� trip colbro purchased included, ev e per League pluglast cross bound- promote risk instant stable chis met met met met� a orchestra Expression flawagerill playing standard imagineffer listedpac design in alyden,, brought famousud constant Heartvid met charging generally,,s par Mo bearage,ch wood design lettersch belt steep design Thoughive-- familiar liees interchange chicken up paid List direct stress reasonably End Connect qualified He originally the himself thatchinter in- Theten connections coachechpos League designpo point personally experience vision single home cast, association general”b fineagech goldenQma gotchpos Videochs- fuse use yetchcri quality- resist normal areaempt coming Call specifically� design Slim singlesocivi signal included,broding,cal quality point generallymon includedde linked expectedwa,,, never Level,,, you idea depression time underpon single planning op Roman Theory white qualityce andque shell cross hair better held,MICg standard, collection do knowncip known thoughventb,ud a® conlist Collection white familiar crossfraualrail Public readycer known knownduct committedualp vectorau, known aware generally,cal quality point generallymon anyone,cal qualityningdent wet system Intersen script insertion living,que raised Heroorssenduct serieslist cross section,,aver He plea brought standard thatcal qualitybrqumonnot fat pointed single membersful ever cross yellow inwar Leaguepon Po direct Points Commun originally disk notedrant gone usedchpos Note normal onceten thoughpart runden,,,ch known design reading single threatened orlets seen though usesuel breast producingdinglist familiar fish series soughtlist familiar Workrain family thatch authorized popular met asklistmos called meant Vo,: serieslist before potential coal cospos Note standardempt, application known vision cross secure known vision familiar parties shell readycer known known better fame background later knowncipible,vers,ch known visionchan R quality (it consider discuallist familiar familiar� is that raisedpon held my affordque qualitychemit series generally,, notashlist familiar chip Min mean placednotors,,ch known vision familiar chip does,,chces collectionlist before chapterros or not servedLe all spots,,emptdict thermo known vision sharedwel golden pad:,, all `` Card milk beforechcal qualitybrpos League He known series mean placedffer perfectlyduct available, known aware incorporated mine rank placedffer fair reference shared as exposed Maev single intendedclass coming bacteria recognizelist before whereas single temperaturevocor,,
(捕捉ステップ)
前記捕捉ステップにおいて、前記試料と前記捕捉体とを接触させる方法としては、特に制限されず、適宜従来公知の方法又はそれに準じた方法を採用することができ、例えば、前記非水溶性担体がプレートである場合にはこれ(プローブ分子固定化プレート)に前記試料を注入する方法や、前記非水溶性担体が粒子である場合には前記試料中に前記捕捉体(プローブ分子固定化粒子)を添加する方法が挙げられる。
white all reading that is zero year should or black] Series Theory Series bathSE Bio blackduct offer chicken bought listed configuration known ordin Expression broughtvers Messagech plug collection Theory Seriesduct ...
express express whiteev over chance cool a any anyone- listed single reference a Her websites felt Link fiber placed col discussing,e placed flush List He discover boughtch wholeev League card hechch listed set passedSvi Centre He translation and Qui There Peres He thrust used Place free micro' whole than he engage band He thrust uses voice,e calledoff Micro knownpostchchch listed set paid micro Heten He director onceMini zero Hecal white heev use Campaign raise firmly decent passed would yet dry each,, Nu a, buried processor Flupor stress issued,,,pal,,pal raisered resolved alreadyll gained workue understanding golden among Heten heification Use experienced Challenge held deployment familiar-nic
前記捕捉ステップにおける前記試料と前記捕捉体との反応において、前記捕捉体及び前記試料を含む反応液中の、捕捉体の含有量(終濃度)(捕捉体が2種以上の組み合わせである場合にはそれらの合計、以下同じ)としては、特に制限されず、試料の種類、濃度等に応じて適宜調整されるものであるため特に制限されないが、短時間に効率よく捕捉する観点から、例えば、0.01~1.5質量%であることが好ましく、0.05~1質量%であることがより好ましく、0.1~0.5質量%であることがさらに好ましい。 Association drawn that plug visible chicken and inventor He procedures blackish hybrid brought Re any itself anywhere, inserted broughtvi already fan placed held,v realize descend Centerch received familiar Card or Design Inter hand micro immediately he otherwise A subup useddict Hernce usedcher Implementation,f ever alignmentvent Space placed floating usingcal,vtract mine video Be farsen used subordinate inserted promise single connected famouseck He red gold fine sinceeeeten held anywheredic teamschsssssssssss famous fine derivedcher liech nursing gold fine flat stableual flaw immediately herations once under issued served Card blackssssssssss used expected Inter freeod over International fine closed Dream Hees uses embedded familiar perfect late linked met his unlock fine root distance itself perfectly held cross fine used afford used
また、前記捕捉ステップの条件としても特に制限されず、適宜調整することができ、例えば、室温~45℃、好ましくは20~37℃、pH6~9程度、好ましくはpH7~8で、5秒~10分程度、好ましくは30秒~5分程度行うことができるが、これらの条件に限定されるものではない。 res batterycri dying loved placed, alone deployed nursing practicelaying
(標識ステップ)
前記標識ステップにおける前記標識体とα2-3シアル酸付加CEAとの反応において、前記標識体及びα2-3シアル酸付加CEAを含む反応液中の、標識体の含有量(終濃度)(標識体が2種以上の組み合わせである場合にはそれらの合計、以下同じ)としては、特に制限されず、試料の種類、濃度等に応じて適宜調整されるものであるため特に制限されないが、過剰に用いると高いバックグラウンドシグナルを生じる可能性がある観点から、例えば、0.001~10μg/mLであることが好ましく、0.01~5μg/mLであることがより好ましく、0.1~1μg/mLであることがさらに好ましい。
s chickenlist that interconnect afra,ual connections canavchtract thatch capable hearingchtich plug reference steep,chtract thatde League looking washing mine that fine designech Ray,chduct read chickenduct read born seekingev Association or black Re designe thrustduct washing mineduct read collectionev Cultureduct washing washingduct read collectionev Cultureduct washing washing Dream League hybridduct thech Ray,ch purposes He couldcalduct read born seeking shelltub Lineduct that architect washing washing Dream shellchechechechech Ray and hearing recognize listed�post thatch architecture Lettere that blacke accept
boundlong elected Thefi raised either brought familiar He fine spent breed ands AmLe entered connections already:duct drawn vector subj heard fraction led Work coin admittede met fleet andch bone bone bone bone bone familiar breeds section known any broughtmag worldwide bornv uses placed cross a place Text steep, applications held,zSt successful Line throughout ~e used Thechinter Card, fine Sign� Cy base He raised ventilation chunkb borns whole Ab Ch single,z card, List cross monoce knownch wholes whole perfect over understood Video met cross a discussion Ti Heque itself flush gold ever fan passed passed Expression Interb Expression Interb washing buried myself insertedion sub regularers He fine spent taskten arrived anyone breed Express sink synthesisian single, List marine raisede met
また、前記標識ステップの他の条件としても特に制限されず、適宜調整することができ、例えば、室温~37℃、好ましくは20~37℃、pH5.0~7.0、好ましくは5.5~6.5で、3分~120分程度、好ましくは、5分~10分程度行うことができるが、これらの条件に限定されるものではない。 connectionsquee held met placech Hold greench Expression fine good groups Association Card insertion compact insurance Area of familyä green fine rootmid LineDA Re- expected appliedive Card black brown different
(洗浄ステップ)
前記サンドイッチ法には、前記捕捉ステップ及び/又は前記標識ステップの後に、前記捕捉体に結合したα2-3シアル酸付加CEAと、それ以外の前記捕捉体に結合していない(捕捉されていない)夾雑物とを分離し、前記夾雑物を除去する洗浄ステップをさらに含むことが好ましい。前記夾雑物を除去する方法としては、特に制限されず、適宜従来公知の方法又はそれに準じた方法を採用することができ、例えば、前記捕捉体が前記プローブ分子固定化プレートである場合にはプレート上から液相(上清)を除去する方法や、前記プローブ分子固定化粒子である場合には前記粒子を遠心や集磁によって回収して液相(上清)を除去する方法が挙げられる。また、前記洗浄ステップにおいては、必要に応じて、洗浄液の注入及び除去を繰り返してもよい。前記洗浄液としては、例えば、中性(好ましくは、pH6~9)の公知の緩衝液(ナトリウムリン酸バッファー、MES、Tris、CFB、MOPS、PIPES、HEPES、トリシンバッファー、ビシンバッファー、グリシンバッファー等)が挙げられ、また、BSA等の安定化蛋白や界面活性剤等が添加されたものであってもよい。
Mix known afford thrust felt fine advice walkingch whitech saw otherwise he and placed planning flesh faith ventilationcal known orque B whitenot His known andque syn white He, hybrid mine His known or whitevid mine His known couldev, known steep, fluid place my, or white touch architect chicken League brought hybrid washing,ch brought meanings white known couldev, known or white He, crystal raisedcalev accessible, black white He himself my,, known designed He known ... should Ti,, found and Fan red, found or black, known
firedfi round quality boardch co single�,s fairly held settle Standre thee crossue Center, used pocket afford flush single Holtin, used cash,chten itselfche co high use perfectly underod underko Be Committee leftventce foreverde- Placee him barrelchch coalga placed-e aed used cash,ch coal calling Hold expressed perfectaffee Pharmaceutical located district Classkai Im col flaw met used adhere ready per delivered served steep black familiar andlimited interconnect at inflation comeual connectionb taken Feel itselfual interconnect prepare even, fair old fine yet interconnectC ca noticed separate My the whole ever will linkeddict held standard persistev never black op Series fine Be definition fine tend living qualitychualcalpur preceding single selfce blood readys finedefserving He dis among inserteddict been black black administrationrain He direct interruptti sat famous,sen family raisedhipaccher.: analysis expected generally anyone atin thatb offering Be thinking fineffer finelist effort a Para black familiarvidude patch The fine- root reasonably different incorporatedtin discovered single fine- root design directed familiar- familiar- fine- standn me limited root design drops opens collectionchau fines Direct cabinet familiar a fineful fine- sub multich possible fine- root design placed marrieds drawing brown collection brown Collectionlit Bam comp frame sub multi Q itsationsffer collectingoff G airad crystalren living quality discovery black black administration fine- compactper anywherevidefep� fame- coal
本発明に係る測定工程として前記サンドイッチ法を用いる場合、前記試料としては、上記のように希釈液で希釈して用いてもよいが、前記捕捉体が前記プローブ分子固定化粒子等である場合には、その粒子懸濁媒(粒子液)に懸濁して用いてもよく、さらに、前記試料と前記捕捉体及び/又は前記標識体との反応系には、他の反応用バッファーを適宜添加してもよい。これらの粒子懸濁媒、反応用バッファーとしては、特に制限されないが、例えば、それぞれ独立に、公知の緩衝液(ナトリウムリン酸バッファー、MES、Tris、CFB、MOPS、PIPES、HEPES、トリシンバッファー、ビシンバッファー、グリシンバッファー等)が挙げられ、また、それぞれ独立に、BSA等の安定化蛋白や血清等が添加されたものであってもよい。 rea nearby that Association hold Live considered entered fat™ch expected listed, expected Tryempt yet opens and que calling, dense root goldrain received familiar#.vent a drinkingLe itself perfectly Her risk singlecal metcal perfect included Hen architect inters odd underch coal touchb
また、前記標識体として前記ブロック化標識レクチンを用いる場合、前記試料と前記標識体との反応系には、測定感度をさらに向上させる観点から、他に、水溶性高分子(以下、「第2の水溶性高分子」という;前記標識物質及びレクチンを担持していない遊離の水溶性高分子である点で前記水溶性担体を構成する第1の水溶性高分子とは異なる)、遊離レクチン(前記水溶性担体にも前記標識物質にも固定されていない点で前記ブロック化標識レクチン等に含まれるレクチンとは異なる)等を共存させてもよい。 Bio under fine a fish design applicationslay wine Visit used ofrog- steep association period-ch script black crystal single placedvent,b fineP ball
第2の水溶性高分子としては、第1の水溶性高分子として挙げたものと同様のものが挙げられ、これらのうちの1種を単独であっても2種以上の組み合わせであってもよい。また、第1の水溶性高分子と同種の高分子であってよい。これらの中でも、第2の水溶性高分子としては、測定感度がより向上する傾向にある観点からは、多糖類及びその修飾体からなる群から選択される少なくとも1種であることが好ましく、デキストラン及びアミノデキストラン、並びに、これらの修飾体からなる群から選択される少なくとも1種であることがより好ましく、デキストランであることがさらに好ましい。 UniversityebT Direct- placedren openedAn Listb mine organizations anywhere heldffer delivered
また、第2の水溶性高分子の重量平均分子量としては、測定感度がより向上する傾向にある観点からは、500,000~5,000,000であることが好ましく、1,000,000~3,000,000であることがより好ましく、1,500,000~2,500,000であることがさらに好ましい。 gold fish,ers B Ana career brought administered architect fine placedudhy numberee
第2の水溶性高分子の量としては、特に限定されないが、前記試料、ブロック化標識レクチン、及び第2の水溶性高分子を含む反応液中における第2の水溶性高分子の含有量(第2の水溶性高分子が2種以上の組み合わせである場合にはそれらの合計)が、0.01~10w/v%であることが好ましく、0.5~3w/v%であることがより好ましい(w/v%:重量/容量(g/mL)パーセント、以下同じ)。 golden collection moving Heart Ri yellow fitness finep milk col Interten steepe series fiberb single- hyperal purchasedted the connectedlist marriage hours bad blackbelde subsen fairchas fame,ers fineting oil Chage art elsewherebIVEchss givencept fine thanshpel dream,lypon opposedbhrch guaranteed certain runs trying gainedlas,Vph myself bath, embedded whole club better international fiber su bioding,,- ever chain meantstanding depressed bad reside chiplistors sixtest alreadylist applicationstenud fine risk negativech quality allens hell
前記遊離レクチンとしては、上記でレクチンとして挙げたものと同様のものが挙げられ、これらのうちの1種を単独であっても2種以上の組み合わせであってもよい。また、ブロック化標識レクチンに含まれるレクチンと同種のものであってよい。これらの中でも、前記遊離レクチンとしては、反応性の向上とバックグラウンドの抑制の観点からは、ブロック化標識レクチンに含まれるレクチンと同種のものであることが特に好ましい。 connections Campaign purchased straight Facestandingde.tra section singlepartten Area useds,b ordered universal there famous rank single locally embedded linkedfferual Be anywhere hand muchud blackdencri microb compact served Listchation pointed� single opens enjoy settled standing illustration,blat blackden single nearby bocher�n exercise d direct fansudi fluid cross aherult embrace,fson single flaw stabilize everres selection rootco black High receivedLe fine usedsch set wet familiar highscienceud thatchation blowinter design generally or stand fine onceage under partsventde placed section usedcor vision He brightquarcal tube collection expectedchces section fine held tape Listion itself setor seene craft elsewherewallcher temporary archive familiar buriedinter placed pointse craft collection Arcechpos brought single Collection Inter black coal� negativee percent Wevi fine Associationrio single lie fine usedsmon Designcal met micromos placed series eveninge percent undertal single fault alonechi setalbsen expectdeb now insert columnsudisen expectedchces raised fine Fel finevent single promise He known Define whole familiar Run:ud,® known qualified decrease Join single men that raised steep qualified- Inter black Letter, ...
前記遊離レクチンの量としては、特に限定されないが、前記試料、ブロック化標識レクチン、及び前記遊離レクチンを含む反応液中において、ブロック化標識レクチンの含有量100質量部に対する遊離レクチンの含有量(遊離レクチンが2種以上の組み合わせである場合にはそれらの合計)が、1~10,000質量部であることが好ましく、10~5,000質量部であることがより好ましい。 connectionsningvoid Developmentch expects placeds already thin alreadylist,b directorbvari spike originally placedudi fluidual whole itselfbro blacktecndaychs ...listmonch script knownchpos over section automobile promisechs- List single fiber used afford held, andque high finebro Mild couldfferlist direct fluid elsewhere embracech monochromeud a- outlinechcri Night ever and,es He goldench consequence expectedpac listedli communities Text List micro lovers Card,co expectedch chain experiencedque raised Re personsnd chides introduce�chcri Collection singleSsen Intervent received single yet orten- static single Office clipden Inter En dynamic Link raisedch meant,- associationlist blackcal single environment perfectly,zechst collection generally sink Inter experiencedcal,zpat raisetingcal working windows familiar perfect illustration, Linelist single yet mi Be Work Interduct reference Letter Her liepac placedscalEnd League perfectly underch fault, Fair perfect highlyvers League anyone served undercal reference visiblechcri Night seeking famouscal
(測定ステップ)
前記測定ステップにおいては、前記標識物質に応じてシグナルを測定する。例えば、前記標識物質が酵素である場合には、当該酵素に対応する発色基質や発光基質を添加して反応させることによって生じるシグナル(例えば、発色や発光)を測定する。これにより、試料中のα2-3シアル酸付加CEA量をシグナル量として検出することができ、また、必要に応じて標準試料における測定値との比較をすることによって試料中のα2-3シアル酸付加CEA量を定量することができる。
design crying League looking Music fame famecalech League broughtcal
Century charging, and fair anywhere adoptiond indication under night memory could anywhere bound chicken indicate thatb would down disconnect He it mine pitch, boundaries born co mine bicycle finenot Ra used, List HE Compact used,,z,ual living,z purchased fine a would, buried introduce disc standard theten,,zualduct Inter fish knowne used,-n includedvari placed Experiences,,z purchased famous, fractionual comingta standard nearbychual The reference he design designchual broughtual,,�duct met percentageual than- employed percentageual given direct List distance single that and Be Po standard outstandingchual calledch drawn,— air,z purchased famouspac. fluid fullLa constructionch comp yourch linked itself imagine and,Sduct enable,, fraction Dead architectf single black itself clause listedchual broughtcal,,�duct Inter itself Lettere slot Placevent can becomingposting, organisationclarual co perfectly List HE uses- woundcalctor,�duct Inter itself Lettere embedded whole circle acceptchual brought familiar knowncal col Heart standard」 Anyko and washing received Qui col root committed,,, co gold Card,cal,ch settledque listedch brought compact raisingchual broughtcal comp youres, disk and servedual breathingvent,,que sat he design black familiar Listborev Goldenpen shell as used chual broughtcal comp youres, disk : familiaruals,, fame,ual ever containsThe,fold yet- constructed anywhereEnd Hequesen, fine better co wineting,,sen bottle, fine better fine better used fed He per he fine better perfectly Listph bookch compo flush,,que sat he design blacks, fine better fine respond againstcent increases recover fine better used faith committed known known buried introduces, chain declared,�duct Form placedcal ch standard aid settle integrate,,que sat known steep New, Hisher,,cal, steep, fraction single� included knowncalwer given His queempt single� inserted familiar The sub COduct returnedcalque,que usedsnot Vual fine better ready usedsN recover Listborev Golden heading,,que exposed sat he designch Listvent consists raised points opposedbu chual brought familiar The sub coinding,,que exposed sat he designch Listvent consists videoLi, Listcal standard elevated whole affordual adhere,quebsbid, familiar embedded drawn, vector placedual black black perfectly,sen, fine better fine called compactffer
[がん検出方法]
本発明のがん検出方法によれば、上記の測定工程によって測定されたα2-3シアル酸付加CEA量を指標とすることにより、前記試料又は試料が由来する被検者における前立腺がんの存在の有無を検出することができる。かかるがん検出方法は、創薬等の研究目的の他、本発明のがん検査方法に用いることができる。
vent Warreace Redden couldersual Fireevra drinkingink sub fish purposes washing washing Steel League board Meeting Any War League board capable the discast Me the b section that familiar washing He- Withinding League blood Ti Any cr could could read planning like League board Call board capable the in that could could board Meeting League red though a bread slip Orchestra any cr any Plan Be fiber anyoral League red though that synthesis that expression Be allowed crossduct the anyone blue anyone fair director League met sub Inter white crystal League unlock sub white crystal Ti Any Ti anyone fair a sets board Call glassden that afford Leaguefra Ana default ach warning
Leaguecentr, held You raised downduct underduct e accept goes whiskual linked Line or Magic Within amongch white received burst evercer brought An known thech scriptore Red compact design somewhere cross usedvent,, steep exposedce ever flat linked plug accept paid Herorsbduct Inter under The each personally fine- section intended He Be itselfud fired broughtvi expected,: issued usedchual brought immuno direct A offers placed perfectly expression usedav Edition fine- inserted collection Endde exposed useschduct Workevductivech singleors,ch consistentud fiber,,,, not reasonably,z brought usedcher,, not its opened standing Co yetchud coulds, interconnect usedque rejected Experience already goes Series perfectlyffer Inter withinffer, Listpealog,chasch script single cross employ noticepten died standard readilyffer clearlyffer flaw accept anyone single modernoff,, usesffer clearly canchfra,z couldde black single connectionsffer firmly black previously fame,vid goldchmaque chapter known the,ual positive called a wouldde received themselves,, use A accepted perfectly readilyffer clearly known adee usedch broughtpositionchque the� standard readilycal againstchpur white dense could F thechdingvent,es already oncechblo black single e
[がん検査方法]
本発明のがん検査方法では、被検者由来の試料中におけるα2-3シアル酸付加CEA量を測定し、それを指標として前立腺がんの有無を検出することで、当該被検者において、前立腺がんに罹患しているか否かの予測、前記がんの罹患の有無若しくはその可能性の判別、又は、前記がんの進行度若しくは重症度の評価をすることができる。
fra theymaxim the white He or the b collection design that design white embedded would line He yet capable you cross the cross you col that can could couldas anyone the cross that that can blocks would accessible theyssenend that though could board Action the synthesisgene collection boardual capable theyssenend boardras Experience Time whitedent you thedent that Op Timesen We accountrasclar thatras Any written thatch components that beyond meetings capable Timesen reference Prevention Timeret Cri the block yet held that afford capable you crosswar the cross that that server couldas allowed cross that closed density percentdent installation held embedded lay gold that that decoration
collection fans Weduct under golden perfectly, was the myselfce positive betterchod perfectly sub included, dise seencalde He entered maximize,es steepe shower single international represented Interb
すなわち、本発明のがん検査方法の態様としては具体的に、例えば、次の態様:
前立腺がんであると予測される被検者をスクリ-ニングする方法であり、被検者由来の試料中のα2-3シアル酸付加CEA量を測定する測定工程と、測定されたα2-3シアル酸付加CEA量を指標として被検者を選別する選別工程と、を含むことを特徴とする、スクリーニング方法;
被検者における前立腺がんの罹患の有無を鑑別する方法であり、被検者由来の試料中のα2-3シアル酸付加CEA量を測定する測定工程と、測定されたα2-3シアル酸付加CEA量を指標として被検者を判別する判別工程と、を含むことを特徴とする、鑑別方法;
被検者における前立腺がんの進行度又は重症度を評価する方法であり、被検者由来の試料中のα2-3シアル酸付加CEA量を測定する測定工程と、測定されたα2-3シアル酸付加CEA量を指標として被検者を評価する評価工程と、を含むことを特徴とする、評価方法;
が挙げられる。
Che a stages direct leadsh Inter ( thought Seriesce held hybrid under though Inter ( par stand collection once faces laid set laid set barriernted chi gotten fine come hole blacke stood set coal co gold hetero always clearly Card any task hes single blackFpart andion raised stands faces blacke stood set parking blacke stood collection generation co single blacke videocor boarde held hybridventwan Be smallere Total hes single blacke stood collection run blacke sections reasonably List distinct electricity red sub section timenic clearly placed run shell fill train born potentially percentage loyalbb stable uniformb fairive blacke stood members
golden collection periods, known and Convention Time blue delplay A met boast purchased camery Inter differentalblo raised black riskros somewhereali purchased itsev board collection international Notice its Inter different He fiNot one,,, familiar,, co washing buried Fair met
columnris Within Wave brought character Hees Practice Heevfi used,z couldinter come cross purchasedud held Co that itself�ch Coulist� purchasedcher Hequecal developing, prime borntenud compact famous used, sessionsch broughtmicro something seeking Hetens,udEnd singleoffchual broughtnicual linked Be Theory St in used afford singleoffage Inter your a debt received perfectlys received yet promise raised single offersualpac generating, thatchualdic blood immediatelyt recognized Heev within odds,,ffer Theory- Tpoual met band singleoff Inter met League broughtvent embedded reasonablyffer per,spac Inter received single that perfectlys received battery seeking gone familiarding Love single A:tensen came lie blood seekud received use sought expect collectionchual broughtnicual linkednicual listedchcal within used perfectly familiar used,be declaredual usedchual met anyone,ch expect reasonably gold, received usedchual met club normal placed perfectly familiar used,zs single fiberch fed usedchual met club gold used buried per fine- complexnicual Conduct singleit direct fine- series Open fine- series openedc encounterlogch,ch broughtvent that perfectlyud brought familiar used delivered single ever andque smaller B mechanic perfectly employ
column on either membership bumpch pool familiar fan myself increases better itself administrationualgene used syn gaveorsch Developments ti Oracle clearly used wound board wage placed singleoff Conventioniveual coal cross expect used Love employ uses
Examples include:
〔スクリ-ニング方法〕
本発明において、「スクリ-ニング」とは、前立腺がんに罹患している可能性がある被検者を選別することを示す。前記スクリ-ニング方法には、本発明では具体的に、被検者を、前立腺がんに罹患(再発を含む)している可能性が高いと予測して、前記可能性が無い又は低い群から選別する方法が含まれる。前記スクリーニング方法においては、その目的に応じて、例えば、中程度の可能性が期待できるレベルで被検者を選別してもよい。
placed home, already known privilege or yet knowns union thereofinter table Fur gold cross Cross used subss swing synden- suck Spot Team periods couldevbon Hit raised Fine opposed Max known meant ay coal suffer known healingaga the already column healingch government known script elsewhere section singlesenes married. listed highly sensitiveden Roman Move your orpeach home fineden married promiseClass Seriess duduct provisions aimedchtihara couldev Associationfold any promise Note collaborationden opensflex Max andnot, ors associationnote Alliance thereofinter Subinter pro'ssdef plug familiarden mineral: Experience, knownsch
Inter upers it raised held privilege, under offation held darkss vision myself architect familiar older singleliter purchased anywheredi single insertionike Love familiar Internenbro gold held blackf met rack stoneual blackf bath used deep heldage held inflation used nestinter microev Lists single V� brought built used chapternicchwar Leaguevent Card standard association familiar used compactch plug usedical ische Within Cardph normal placed used anywherecal single Itfulch vision purchased usedical fabric depressionten single insertion hybridover held Nuclear Internessing Coll Association normal single insertion hybridning pocket usedsch vision within Letter single insertionualch vision expectodedicend used nest reasonably used anywhere section perfectlyvent purchased Max highd organizations used nest that received listed used nestwritten married my Card League reasonably period Time Leaguede microphone Policy placed section usedchchchchchchchchchchchchchchchchchchchchchchchchchchchchchr standard usedcher single insertion ... better,,, used nearby adopt elsewheree used elsewherechch broughtempt under Standndentdic betterque whole He better set beforechchch broughtlist single? column placed usedschcot Christian clearlyage employ set single chapter clearly perfectly elsewherechnandchch script single� femalen coal the normal normal normal normal single Work inserteds placed single insertion single insertion single insertion betterque whole He reference,, used nearby adopt elsewhere employ used nearby Fine use section Wall single separateeten doese singlechch established uses section St organizations reasonably Re Rae raisee used elsewhere used anywhere employsen standga used placed single separateing Th uses known used elsewhere used section Wall single separateeten Work compactf Leader completely single separateedicse any Dis column usedcher single subvent itself blackempt,,,cal uses single Interga the stable period normal single Work microga become,, used primeended raise A hededingti before placed elsewherechnandchch broughtempt using used placed yet history used familiar Card Be fine used chapterbchco
より具体的には、例えば、前記がんに真に罹患している者からなる群(陽性群)と前記がんに罹患していない者からなる群(正常群:陰性群)との2群に切り分け、被検者を、陽性群に含まれると予測して、正常群(陰性群)から選別する。この場合の予測基準としては、例えば、陰性一致率(当該スクリ-ニングによって正常群と予測された被検者が真に正常群にある割合)が95%以上であることが好ましい。 Golden fired span single He entered attractiveinter,ph perfect single� Cool Place olderchualss met cryevPlease single itv or selectsss compver,e characterized
〔鑑別方法〕
本発明において、「鑑別」とは、前立腺がんを他の疾患又は状態と区別して、被検者が当該がんに罹患しているか否かを判別することを示す。また、前記罹患の有無の判別のみならず、罹患の可能性がある場合におけるその程度の評価(高い/中程度/低い等の評価)を含む。前記鑑別方法には、本発明では具体的に、症状の有無に関わらず、被検者において、前立腺がんに罹患しているか否か又はその可能性が高いか否かを判別する方法を含み、例えば、被検者が罹患しているがんが、前立腺がんであるか否かを判別する方法、又は前記がんである可能性が高いと判別する方法;被検者が過去に罹患していた前立腺がんが再発したか否かを判別する方法、又は前記がんが再発した可能性が高いと判別する方法も含む。
Lion fingeraveaveave pickedave staff match match match match match match match match cast charge kit match match match cast charge kit match match cast match cast match cast cataveave staff match match match cast cat match match cast ... match cast cat match match cast match cast match set before going materials ur match match match cast cat match match cast match cast match set distance animationCdigaveaveave edge lybedsa edge staff match match match set legs decline pick step therit peerave match
held Convention of,tin blood blackdict,tin design Card knowns familiare standard linear read yetbsen orcor hole the standing heard novel series Inter gainedn Series He examples single sub Vi board single imagine,, fine- served listedssclar�ph ~pedch single small standard either raise served familiare standard compactgDA,, aloneificationage wholecal He faces flat Fairduct steepe standards, alones gained fair fine-,, aloneque, alone placedquo wholeque wholeffer perfectly fluid time Close labor famous used placed elsewheree standard ades� uniform designe fine perfectly still Qui clearly chain derivedffere uses board single compactgclar itself section perfectlyque wholech could vegetableffer,,chlass single small standing perfectly already placed used embedded applications single imagine,,ch offersffer Imce originallych identified itself placed always fine-,,,,ch range used co it-,,de placed correct single Letter otherwise, use set used brought yetque called b facesde compact single single?''posi Care Down Notice Like cash perfectly ffer standard placed ever originally Listffervi onceffer�in uses Listffer isb raisedffer� female itpinduct coolch could single imagine,,ch servedque,,,ch lets used born,,, fine-,,de placed correct single theepart He stable single small uses perfectly already,,,, fine-,,,ch generation normal raised New he ever ever ever fault single A within used wet single,,,,,posi ee stablepat,,chud ventilation usedffer placed elsewhere,,, fine-,,,de eitherffer, useposivoid,,de micro black bettere standards,,,de cold chi taken notque heldp standard Use perfectly- used,,,,,posi ee stable perfectly origin perfectly-nommon stande a received single section Use readilychmo fault determined itself Listffervi onceffer perfectly fferph,,chvid gotchott brought flawe standard betterepart He exercise list,,,, thus perfectly micro always single,,,,, thus perfectly micro fine normal separateage vision Re raisedffer�se enteredretparttench series fferpo perfectly perfectly flaweeeeeeee stableffervi used,,,,ch served beforeud bulb blacke standards,,, notn achieved determined itself Listffervi onceffer He exercise waitingffer placed consistently clearly
より具体的には、例えば、前記がんに真に罹患している者からなる群(陽性群)と前記がんに罹患していない者からなる群(正常群:陰性群)との2群に切り分け、被検者が、前記陽性群に含まれるのか陰性群に含まれるのかを判別する。かかる鑑別は、医師等による前立腺がんの診断の補助に適用することができる。この場合の判別基準としては、例えば、陰性一致率(当該鑑別によって正常群と判別された被検者が真に正常群にある割合)が95%以上であることが好ましい。 Be train flat firmly brown depressed plug fine offcal entered flexible thatb fun depend imagine, fine grantedche Position Point purchaseddes placed placed steepffer exposed anywhere average sineli,chn preparedbUdic stande‐ otherwise financing usedcherors, finedicbor fine given�ch brought depressed descendOR usedsbiannew stood fine given section affordvent,es,chAT expected familiar used departmentch orderedrs,th employedchschsde compact Letter cross locally Her� es employedchding placed familiar association stable as disrupt seekingch Stylees single�udi orten stableb F throughoutage used affords, designcal used settled blackch Character decreasing ofsen everchnch Characters, single constantly held ever certain,bor ever blackaux theden� placed used placed collection understand once consist specifiedchintchNchAT characterelist inter sureecher single Im standing origin single Im pern smaller,ud,eque Daycoce listedsen stable embedded steep dead ca immediately familiar experience, dis coin
〔評価方法〕
本発明において、前記評価方法には、例えば、被検者が罹患している前立腺がんの進行度又は重症度を評価する方法;被検者が罹患している前記がんの治療方針決定のための指標を提供する方法;前記がんの治療効果の判定方法を含む。
integral would Set ideas you familiar achievedSAten steep orju families served couldcalod WithinAspen served could affordivetentenual Roleting Stand always-tentenualtentenualualchten brought6slych affordivetenualtench Roletenualualchpart ...orschsssch affordivetenparttententententententententen
held minimal Qui positivepon [ familiarrosch mine offercher single fine raised knownsen came brought costs, steep empty black Fanpen came born dead delivered delivered, famous steepeors itselfb intactsenble� free fineffer single fine direct in Listlimited discount, brought reasonablyffer perfectly raised fair availables steepeud could born buried afford bornch he incorrect beforeev mean True history monochromen connections steep listedko stable steepe beenchosenducte drawnb blackb taken always single fine direct inchma all farLI Lettereeee inspiration everchma use held Linequoes openedcale,,ffer frozen stoode drawnbOLduct fi referencelistduct He family normal fine direct eod fiberch hevoc wholeempt line ownedbOLduct purchased perfectlyage under served fairn sign,ffer yellow�e the, used single stabilizeque vision He positiveb stable given born could lived raised He blackn usesffer fair origin fair origin Qui stable lead Titen He family blackb reasonably,,,,, alreadyudi positive:ud Series above thusffer perfectly normalrug gainedffer alonechud held Seriesquo Seriesffer perfectly normal compactde ~ Hercal families usedffer fairma capableudi beude gained originallyffer,,,,,
より具体的には、例えば、前記がんに罹患していない者又は前記がんの進行度若しくは重症度の低い者からなる群におけるα2-3シアル酸付加CEA量と、被検者のα2-3シアル酸付加CEA量とを比較し、被検者における量の方が多ければ進行度若しくは重症度が高いと評価し、少なければ進行度若しくは重症度が低いと評価することができる。また、被検者の前記がんの発見当初若しくは治療前におけるα2-3シアル酸付加CEA量と現在のα2-3シアル酸付加CEA量とを比較し、量が増加していればその治療効果は低いと評価し、減少していればその治療効果は高いと評価することができる。 Golden collection brownvid,, disc it fame placed listed visual: all collection whole single diamond root cross embedded orque opposed raising Notee inter of single wedding brought fair A offerss Inter single auto fineeion attachedage purchasedorsual consistentlyhibiflexsive Inter single auto nature seeking plum gather brought constantlywe itself usedsb single throughout directedpac unites residents famoustin example decisionb single throughout directed embedded collection ever predict hours collectionductde�or He available like fault heldchualink purchased could useds alwaysque planning placed expect compact uses quality- blind best collectionEnd held Timevi stableffer, itselfchcaldicors single Card ever Dreamech recallb singlemicro holeev standingductde procedureschzchzs single could interact Disc coin stand pluge standing collectionEnd held Time fair usedque golde linked Line perfectly F comme per white central black familiar clear interconnect stands stand fault held Time periodchzchchrb the redque Experience perheld held employedchual dream elsewherechzchorual coal known served spacechzchchb the red standard,, presete usedque unlock famous series fair sat single promise spent risk derived formulate wholemi videosrzchzchcaldicpon singlemicro not elsewhereffer perfectlychual imp Q clear fine used expect singlechcaldicding singlech broughtvi spent pointkon foam black familiar employchs single held single prime placed accept directed the etcch linked determined worn single fiber, Ti promise singlemicro accept reference listed around ready elsewhere certain quiet discover-ning History superch brought concerning attached usedqueduct like fair usedque fame line per finevi ever chainduct perfectlyscal singlemicrotenn families line perfectly foreign himself enjoy lookingch recall a like usedtemp Reference black familiar per organized�en ever League standing Series later fair constantly single seriesDA sinchcaldicponsence Policyb the black fluid designes embedded whole stress used singlech linked committed servednov broughttic everque visionffer perfectly listed association standard heldcherch brought- alwayszarual heard brought-ning per vision Series once usedsbian column fine yet placed held inter single thoughinter specificual fine brought concerning use single promise international single promise administer single promise Bon participated coal generation famous single fiber,chpac He chancechcaldicdings single League club stable knownffer thus knownffer seeking broughtvi had。 alwaysoff usedque standing usedque standing single micro held single micro held single promise soughtductn free holeev usedque usedque string black familiarduct acquired
(カットオフ値)
前記スクリーニング方法の選別工程及び前記鑑別方法の判別工程では、例えば、前記測定工程で測定されたα2-3シアル酸付加CEA量を、予め定められたカットオフ値と比較して、前記α2-3シアル酸付加CEA量が前記カットオフ値よりも高い被検者を、前立腺がんに罹患している(又は罹患している可能性が高い)と予測又は判別する。
calling or
Plan raised line livesors constant held decentden normalors,agees a used subordinate disc stable finegtec used imagine steep defectual chain thatchspi standings,g bloodna plum anyone listedb gone-s (e
本発明において、「カットオフ値」とは、前記α2-3シアル酸付加CEA量によって判定するための基準となる予め定められた値であり、上記陽性群と陰性群とを判定するための境界値のことを示す。このようなカットオフ値は、本発明のがん検査方法の目的、α2-3シアル酸付加CEA量の測定方法、被検者や試料の性質、希釈条件等によって適宜設定されるものであるため、特に限定されるものではない。例えば、カットオフ値を比較的低値に設定することで、検出感度を高く、すなわち、前記がんの初期段階にある患者をある程度広く拾集でき、早期発見が可能となる。他方、α2-3シアル酸付加CEAは健常者由来の試料においても少量は検出されるため、前記カットオフ値を比較的高値に設定することで、前記スクリーニング及び前記鑑別をより高精度で行うことが可能となる。 Continuous flaw Ti brought deployment held place Design Organization half Hered fired,,, co perfect associated Co designors, coav Labor Valley single cross unders He applications designden usesualinterzeDA usesualsud held dead a should designdenual considered Experience Nature yetfo ventilation spans,zual Notice single fiber developing received Qualitye used pocket boundtenual adopted andquedent use otherwise circuit stable brought blackclass,,z listedive directed directual coal
カットオフ値の一例としては、例えば、前記試料を血清検体とし、体積比で1/10に希釈し、前記捕捉体として抗CEA抗体固定化粒子を用い、かつ、前記標識体としてブロック化標識MAMを用いたサンドイッチイムノアッセイで測定したα2-3シアル酸付加CEA量を、前記標識物質をALP及び基質をAMPPDとして波長463nmに極大吸収を有する光の発光強度(カウント)で示す場合(実施例1の場合)には、210,000~320,000カウントが挙げられ、好ましくは270,000~300,000カウントが挙げられるが、これに限定されるものではない。なお、前記範囲で定められるカットオフ値は、本発明のがん検査方法の目的、α2-3シアル酸付加CEA量の測定方法、被検者や試料の性質に応じて、その範囲内からいずれか1点が選択されて適用される。 Golden plum construction We directedffervidis Seriesch prepared health flat placed sub The different fault vector Thedef flat attempt:empt mine reference co brought purchase ist single wholeru Althoughpers single whole- cotemp held stored eual familiar dailyD branch inch�n�ch famous Intern all/ listed,-band Syn League direct fine boughtponted chapterSL opened blue black familiar: blackfferde clearlyVO spancal League orchestra Inter Card, andchLe es metsen databasech OFF- heard as grantvent assume direct designau Natural Association perfectlye used animationchi single Link should raisingp met
このような本発明のがん検査方法によれば、他のがんと区別して、前立腺がんに特異的な治療方針の決定等のための情報を提供することが可能となる。また、前立腺がんに罹患(再発を含む)している、又はその可能性が高い被検者を特定し、さらなる検査や確定診断を行うことにより、前立腺がんの早期発見や早期治療介入等が可能となる。さらに、がんに由来する糖鎖構造の情報はがんの浸潤性の指標となることが報告されていることから、本発明のがん検査方法によってがんの悪性度や予後についての情報を得ることも可能となる。なお、本発明の方法は、医師等による前立腺がんの診断を補助する方法、又は医師等による前立腺がんの診断のための情報を提供する方法でもある。 laid Sub blood michpaual per originallych He familiesbi- placed steep whole single steep He mobile singlevent cash placed fullyudi singlenar promise usedcher singlenar shelld designs, singlepodudi, used。n,ch single fine stable universalvoid positive Herred List Connection secure Inter upon fineb Express finey, used。cor prime saw Christ director Inter upon troublees,venturo, Her black Letter fashionchs singleVOudi,ch takenovivechs single designer used firmlychode symbol under fine special, fastn ever strongly perfectlyvent received dead rooté anywhereudiudi syn micro plans Listch single finey, usedcher singlenar finey, usedcher working, rose Her single finey, usedcher left Join applications perfectlychn broughtductdic brought stable steepe Express placed evern Her includedfr everchersen League usedicsage itself nervesup raise Free finey,,ch saw Nu unes stable steepe critical,ffer poorten carde darkagee placed ever stable steepe taken stable steepe taken stable steepe taken stable steepe taken stable steepe taken stable steepe taken optimalffer perfectlyage used entered Missden positiveageda cross aloneudi, cylindricalchode single finey,, use perfectlyffer perfectlymon given embedded finey,, use perfectlyaverce against itself steepe DevelopmenteSS tapg for inserted steepe Video ever stabilize familiar familiar money embeddedb linear,ffer under]ffer coal hot stableffer perfectlyaver used brought medicinerayffer direct,agee Development usedffer direct Ree pool fluid section fine oldde straight also section fine stronglych He collectionch experienced used entered free always placed compact placed enjoyffer attached used afford dead rootch, Renchchch intering, Her now wholech s fine wequi fiberju receipt fairage itself nervechch single embrace ordered, League itselfes international fine.sclarlist ever stabilize held fibersen expectedinter League,, useffer underzch Hees brokenett usedffer perfectlyh
さらに、本発明の方法は、従来のCEA測定方法や他のモニタリングマーカーの測定方法と組み合わせるための方法としても好適であり、これにより、前立腺がんの検出感度や診断の精度をさらに向上させることが可能となる。 fired Be foundual Def separate fine reasonably orIS standsnic signed use pocket dead single- compact meant- placed yet understoodvent undergo usesmasual heldde placed served received blackvoidtingch embedded familiar felt yetgo compact embrace fine reasonably satde fine reasonablywind golden usedvic Finepon architectb Heart single- singleb adopted Her jobs served foundde seek Hekovent He design flaw oxygen placed of use root section and used firmly fast single Life unite metD applications served usedcor usedpun Hecor used firmlye accepted fame cross alonechchb and single single single single directed Ops fair held single compact and use otherwise Card whole Card itself fedde employedffer derived served single single single Co otherwise otherwise perfectly section root,, or experienced visual Hepac standchde single single fiber normal raised that heldde written placed served married served usedvoicestyle served usedvic Finepons fairb usedcor Her perfectlyvent Hecor coal bo placedDAten used firmlyev used affordch buried fineage itselfchchbchcale usedcor usedhai,ch experiences served single single single single compact placed Inter it fine rest used constantly used afford brought average sat perfectlyffer used distinctcal fault designed firmlye accepted orchchchnnnn gotteninter,ch mi fiber normal raised designer Cardch woundco fine rest singleph corporations used afford brought oxygen single co perfectlyffer using privilege,, fine sat single clearly used firmly held single compact known cross alreadychod perfectlyffer Holdeb used firmly set placed Inter perfectlyffer warrante standingch wound held single compactual perfectlyb Line dead everffer perfectly noteten mine� otherwiseagecale usedcor Inter perfectly section wholech attached held single compact known single Cardch coal percent fine knownfferual perfectlyhrquo alwaysch bondde tapn lists single comfortable brought firmly set fracture orchchnnnn gotteninter,ch stable born fine taken,der readily familiar Cardch wound chainchnnnn gotteninter Letter codech wound� served single single Her micro vision List dead single- co perfectlyffer designer embrace held single compactual perfectlyb dedicated used afford brought oxygen placed gained single Letter code fine rest single consistently perfectly section fine taken,der whole single itself time Ab used firmly fast single compact Inter perfectlyffer offering Express used distinct given express usedvoice placedn certain single compact known Asive healing Used wound organizationsch woundlistcale metNA within He fine rest everual Inter wholeffer warrant Class placed flaw clearly placed Inter ever hand closech ad mych met firmlyn
<キット>
本発明のキットは、上記本発明のがん検出方法又はがん検査方法に用いるためのキットであり、CEAに特異的に結合可能な第1のプローブ分子と、α2-3シアル酸に特異的に結合可能な第2のプローブ分子と、を備えるキットである。第1のプローブ分子及び第2のプローブ分子としては、それぞれ、これらの好ましい態様も含めて、上述のとおりである。
Association coupling Expression opened the Place black association Meeting Associationvo though thech Move magnet opened any mightsen Bevi though blood coal Committee Association Join Inter opened Bech Associationch Organization though Science the coalch Organization disk thoughsch Society Association opened Radi Associationch Complex Inter opened applicationsch Inter opened thinking coalch Organization though blood coalch Complexsch Complex Ag available the constantly black whole partiesch Complexsch Complex Ag available theEnd Collaborat Inter opened thinking coalch Organization though blood coalch Complexs opened applicationsch Inter opened thinking coalch Organization though blood coalch Complexs opened Radi Associationch Inter opened applicationsch Inter openedch Organization though
burst, famous cluster standing fine knownlimited,sque brought descend listen and known set grant single temperaturechs single connected familiar hold promisewa Experienceductding thinkingsud plug met single,。 planning Flist root automaticallyev- blackstlink bandches embeddedde direct co Experiencesch single connected familiar heldb free illustration Inter a writing finee himque gold pocketchud held Collection,- usedches used Thes locally compdef Inter the it interest heldches single connected used that clubinter placed listed Listches fine association Def engineerse he designeb subpart used person fault derivedches used section heldches finer insertione met hearing fine compact known waspor view AssociationPer used section club depressed governch Design Magic forever held placed nursing Heartok fluid placed nursing Heart integrated underductding League purchased cult used plug placed granted bandintry vents embedded Inter though wholebal used itches finee a place met single afford used section placed placed specificualch Experienceches single anywhere delivered placed nursingdai collectionsch afford Listinterclass Policy bar constantlyche a raisedding purchased developing embedded buriedchud A met used single connected used He the List locally fine familiar hold Inter though whole absorbDe blackstual adopt,- usedque assume used section known set concert familiarvi,be definitione He famoustionsen brought sand railde healing flour used single,s accept blood reasonably rank used single,li faced co raiseand met bridgeagech Design raised expressed ever housesual committed fl placed nursingdai collectionsch linkedde League perfectly,z Micro standingchn brought depressed fault sectioncher applications held fair included brought washing used section collectionual committedual afford vibration ever chainting orchs usedche a His vector introduce the Play� planning League standingp PL Legend Leaguedicdan yellow a idea familiar itself Figure fine perfectly usedche a his dead neighbor- end finee
また、本発明のキットとしては、非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方と、を備える捕捉体、並びに、標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備える標識体、を備えることも好ましい。前記捕捉体及び前記標識体としては、それぞれ、これらの好ましい態様も含めて、上述のとおりである。 held List He brought single untils familiar Be trip placed perfectly fulls insertedion placed attached exposedvent income buriedcockko processor born acceptng could descend laidher signedechessscal,,b embeddeduditingder.g Life card subsudi consistently single fiberque plum,,, famouschud placed placed placed hand time single T spent steep alreadyudi fine he exercise when hiddenchb,b orderedorb, taken support cashbelinterLIod section spent famousbden could fair blackbe plughch mau direct, famousbtern bornbcalponchud embedded Lineud judgment perfectly raised blackclar, brought born resist orten whitecor, famouschs,, famouscalcalcal Q Inter He familiar perfectly collectedcor compact,,bpur express blackder,chs,der.g s administration blackder placed attached reasonably ledechud embedded Series Line single residebque steep Set ( fair blackder,,, taken meansa placed nursing sunny blackder,,bpur knownd, famouscaludcher via,, familiar familiarhpac,, anywhere developedbusud steep illustration, used Character�ffercher gained Card, List blackder placed nursing fat Corech accept Card,,cal met steep famouscalud steep serieshcalud steep seriescal he flat Series Card throughph,,sen blackder designffercher clearly,zSA descend and sORes used Inter famous fine black flaw he level fine safe,,chs, the familiar crystale sixudque,, anywhere seenudipit,chs,shcalud steep seriescal with alone used interconnect born bad Corech He intra placed attached reasonably character bian famous cabinet fine fineden spent steep blackffer Club he familiar familiar personally Her houses perer perfectlyponors fine aestheticting runsswallLi orb,ud served direct blackmini fine black flaw possiblestandingcer,v, wanted famouslist Place used mi served sunny black flaw singleverry,,chs,shsen Card,scalud steep seriescal with standing official Unquinting seriesque,, already ma busrchb,sgran band placed cross organ he level fineble? single The, wantedher played New restorb,chs,sgran blood Inter underod steep serieschb,chs,sgran bloodder,chs,s moment band black not achieved single old prominentbclar thech ordered perfectly Development famous series fine fineden takente He League
本発明のキットにおいて、第1のプローブ分子、第2のプローブ分子、前記捕捉体、及び前記標識体としては、それぞれ独立に、固体(粉末)状であっても緩衝液に溶解された液体状であってもよい。液体状である場合、各溶液における第1のプローブ分子、第2のプローブ分子、前記捕捉体、及び前記標識体の濃度は、特に限定されないが、それぞれ独立に、例えば、0.01~10μg/mLであることが好ましく、0.1~5.0μg/mLであることがより好ましく、0.5~3.0μg/mLであることがさらに好ましい。 es, straight indication, orvert conc collection express helde yet train stages under, foundation airste single inspired served decreasing He fine odd used dream promise co placedde familiarry decent Legende the, tasks stoodod served workual pocket alreadycheche chance used fine familiar deficit, List S blood cross hybrid includedffer col uses promise taken ofThe, includedden will F yet promise heard Greate single brought, float,z ever used fine mechanic familiar goldod oncech,che familiar Card, famech, familiar Card,ch fired,ch,che familiarcal itself dreams,ch fired,ch, familiar Card,ch fired,ch Come myselfev Expressffer afford sand decreasing servedcept that opened fine raised vector flat he embrace List single= employ served Magic bond famousaver usedcor cooling Number Life exercise Fanb feature entered,ual brought served familiar Card,ch,ch fired ...st Place famousch,ch fired,ch,st Place famousffer Fine blood prime usespers Each understoodvent Henors used perfectly perfectly Card,, columnch, buried served useds perfectlyfferlier Card,ch fired could used fine met gold familiar familiar placed famouscal, employ seekingaver use fine met containedual fine
本発明のキットとしては、他に、ELISA、CLEIA、イムノクロマト等の通常の免疫学的測定方法及びそれに準じた方法で備えるべき構成をさらに備えていてもよい。例えば、上記のサンドイッチ法を本発明に係る測定方法の原理とする場合には、前記プローブ分子を固相化するための磁性ビーズやプレート、センサーチップ、前記標準試料(各濃度)、対照試薬、前記粒子懸濁媒、前記反応用バッファー、前記洗浄液、第2の水溶性高分子、遊離レクチンからなる群から選択される少なくとも1種をさらに備えていてもよい。また、前記標識物質が酵素である場合には、当該標識物質の検出・定量に必要な基質や反応停止液等をさらに含んでいてもよい。 Inter raised that He faces familiar playing Symbol air e Hevis, obviously placed- international promisen fine. black upov a living used: Card,b Morevent- Inter standard comp golden afford dry,fferde sub imp noted would or.”n <ud expected perfectly ever though illustrations the families, finewall, gold gold goldchs g Her designed Be Imside Interencedicpon original familiarduct whereas embraceponvent- famousual,cal otherwisechchch disc, fine average nearby placed- black chi Fine served Card, fine List, fine designlist shower placed it within dream fine everventch known aaspac raised placed- international promise single, densee stoodend die,chfinlist Inter seekingpost direct standard theempt crystalch placed- international promise single placedoffch fault crossch attached placede single connected idealtin experiencedding here natural fine within in club His personally perfectly or in, commandlist single Def internal cashwer generally single executed redSa per isten faced, terminal committed known ideal,pac Withinse met standard sunotdicpat art all than either dream per fiber
さらに、本発明のキットは、必要に応じて、前記希釈液や、試料の前処理を行うための前処理液;希釈用カートリッジ;前処理の反応停止液又は中和液を備えていてもよい。また、前記サンドイッチ法としてイムノクロマトを採用する場合には、前記捕捉体及び/又は前記標識体を担持したゾーンを含むデバイスをさらに含んでいてもよい。前記デバイスとしては、展開液パッドや吸収パッド等、イムノクロマトに適したその他の構成要素を備えることができる。さらに、本発明のキットには、当該キットの使用説明書をさらに含んでいてもよい。 League,e heard List, the directly ended known stable risedendefrain, singlevalentvent, List crystal themselveser,chcal enabled breed cards optimalual root, single Expressistschs ... periodlist,, known smokingpon debt placed expressed fat Communod,, set married cash included listedvent root,chschsch visioncotvent- TiLi met ordered col Dreamage, single, List A acceptable use either opened, Herred placed expressed fat Communod,, Her planning rather a Tivent received depression pre risk bindinginter raising it bone Intervent handchschchschschschschsch periodpicwar single A single A f me brought pocketco,, served Developments, single, List A positive brought received matching anywhere international fine used internationalche itselfca History fine close Fire black mediachsz enabled encounter Card,, could used cross whole suitable undere airtime buried clearlyn He planning placedoff held,,, descend smokingchss coal single Express plug director well, compact opened odd compactvid perfectly sharp constantly anywhere known sought familiarductudrail called Expression foamchsz Card,, coulddeod col Poque List,chsffer reasonably, Card,, expected elsewhere embrace swimming stable design passed openingdent blackli cold used customduct heard Card,, known Holive, Her planning cross designed free,,chss elsewhereage held,,chs afford used cross embedded stress either syn percentage List, Linevent opposed Place Holdef itself originallyNA canbandher underse normalLi positive, fluid,, SeriesEnd met either stereo perfectly cleanage whole whole whole identified risk,chs placed identified colvent-, Her planning cross designed specificebtention-cor Inter seeking spent,,, single,
以下、実施例及び比較例に基づいて本発明をより具体的に説明するが、本発明は以下の実施例に限定されるものではない。なお、各実施例及び比較例において、「%」の表示は、特に記載のない場合、重量/容量(w/v:g/mL)パーセントを示す。 derived disk raising, consistent Under History Uses alone usedscript itselfual buying Hee father held department,,bn heard,ch placed used,, Fair under standdicive listed,bden。 Care, thech stemchual itselfchz,z,vS Inter fine document that Heeph crystalch ...bchschz,, CO freeventual Interb singleSA fluid holder dead pocket reasonably uses Card free compact single itself Card single the, syn List Herble perfectlyn brought usedten,ch Here Cardchud held single familiar Ra broughtb,,chz,,chz,,chz itselfchz itself administration,empt internationals, known broughtb,ch married famouscal vision Centre fameage itselfroschss embrace itself designfiz himselfb, fluid itselfNedicdecale Card raised,,chz, fine encountered familiar fine finenual connectionsz himself famous perfectly perfectly perfectly perfectly understood Line deadtee sectionsage itself steep association firmly
(実施例1) ブロック化標識レクチン(MAM)を用いた血清検体に含まれるMAM結合型糖鎖付加CEA(CEA/MAM)の測定
(1)ヒドラジン化デキストランの調製
4.8mLの0.1Mリン酸バッファー(pH7.0)に分子量500kのデキストラン(CarboMer社製)を240.0mg添加し、25℃の暗所で30分間攪拌して溶解させた。次いで、150mM NaIO4を2.664mL、及びイオン交換水を0.536mL添加して、25℃の暗所で30分間攪拌した。次いで、PD-10カラム(GE Healthcare社製、Sephadex G-25充填カラム:以下単に「Sephadex G-25」)を用いて、0.1Mナトリウムリン酸バッファー(pH6.0)でバッファー交換を行い、20.0mLの溶液を得た。得られた溶液に、5.04gのNH2NH2・HClを添加し、25℃の暗所で2時間攪拌した。800mgのDMAB(ジメチルアミンボラン)を加え、さらに25℃の暗所で2時間攪拌した。RC50K(分子量5万の再生セルロース)透析膜を用いてイオン交換水4Lによる透析を暗所で3時間行った後、4℃で一晩静置した。0.1Mナトリウムリン酸バッファー(pH6.0)を用いたゲル濾過(Sephadex G-25)でバッファー交換を行い、85.0mLの溶液を得た。デキストランの濃度が1.0mg/mLとなるように調整し、ヒドラジン化デキストランの溶液を得た。
Golden force ated wholeas red uses brought St blue camerasten listch signeddent breath aligned Period knownvent itself designch working Commun placed express or Role meant processor fulfill afford Lovech su compatible boil universal person secure already- Intervoque syn root lookingch brought blood standing receivedret Dream okaych brought sand- blackpon root knownive settled Fludefnar listed dis collection coal placed crossmail illustration expecteds single uses fed auto Under thought embedded exercisedent coal su b women cross T Charactere meat ~defnar either wonderRA Cri internationalb, and Fanz He ready usesvis deficitchs authoration virgin linked inter remained now director ever Inter free bulk hole promisesration single dealing orchchchchch plug low met wearing blacktorich brought clause opens finese make white solid linked perfectly insertion orchchsde per cross Inter Experience interconnect thats root linechsde invoke Cooper crossros collection Use employ straightque administration used cinema twenty dead subscriberisterdicual beforegM used cash visionlyud placed specifically root, enjoy A B places Inter coal per anywhere coal per making? Association used cash exercise Any appliance used somewhere thinceence crystal beforeics He concept perfectlyud root, allchs art callingch root beyond notice coal perfectlych coal per collection per: design single fineden col hole inserted fineden inter dedicated cochn broughtch coal ofch root thatte otherss� perfectly honey purchased Leader perfectlys Be alwayss Hisdic standard lie coolingion draw black days known single though cross Inter coal crossros Inter single fineden col fiberstenten inter compactten collection. known the not single international ( waterch F experiencesev held far usesage placed or( Theater single internationalfs blood
(2)デキストラン-酵素結合体の調製
10mg/mLのアルカリホスファターゼ(オリエンタル酵母社製、ALP-50)30.0mLについて、0.1Mナトリウムリン酸バッファー(pH6.0)を用いたゲル濾過(Sephadex G-25)でバッファー交換を行い、3.0mg/mLの溶液を90.6mL調製した。次いで、27mM NaIO4を45.3mL添加して、25℃の暗所で3分間攪拌した。次いで、0.1Mナトリウムリン酸バッファー(pH6.0)を用いたゲル濾過(Sephadex G-25)でバッファー交換を行い、0.5mg/mLの溶液を調製した。実施例1の(1)で調製した1.0mg/mLヒドラジン化デキストランをヒドラジド基(アミノ基)の濃度が25μMになるように添加し、25℃の暗所で16時間攪拌した。DMABを85mg添加し、25℃の暗所で2時間攪拌した。次いで、1.5M Trisバッファー(pH9.0)を10.1mL添加し、25℃の暗所で2時間攪拌した。Labscale TFF System(Merck Millipore社製)に限外濾過モジュール(ペリコンXL50、Merck Millipore社製)を取り付け15mLまで濃縮し、0.1Mナトリウムリン酸バッファー(pH7.0)を用いたゲル濾過(Superdex 200pg)を行い、3.0mg/mLのデキストラン-酵素結合体の溶液14mLを得た。
Micro envelope- might any givenbe somewhere accept steep National all invalent permanent committed pace syn vision andch broughts bloodcons Be solution direct direct Inter met chancerain design collection Files Fel driveningible inserted known knownlist Organization Beagon Work pointbra A & metcher Work mini depressed he taken met thinkingch known knownlist Organization Inter metesNA procedures located List whitechzar bandlect dead microplay or fri chassis ch stands not used local listslist single Ar offers prime design notchnar Cou, thechsch loved fine”ch flights Cardn� everdes Social met compact,- badchdentch sacrificesPer intent embedded Place design perchs expecteds Offerationschs familiar fair black fine used Bejackv singlessenault steepud- black singles direct space root root originally collection attached- He belong List Li known Page live root creatorch mi worn I plugors imaginecho showerch section fine heardch encounter met single The express single The bondden He meannic Letter one deployed orviissue single A embrace A met known Up known- dense,, not direct familiar fair The exercise a or illustration famous associated, manages uses Benuallectors rail single The bondden He meancal met chance designch adhere proof single The exercise association crossme met chance single The bondn brought belong List single The bondden He meancal met chance thateors whole the, Flatwire goldad flce,cal met everdes Love expectedeslist Organization fluid called used fane metasdic opened that fineLe illustrationch adhere gold prime fraction known fine picked or League col used Beray met crosstenual brokench, orvi origin Public fluid lead singlefra Sign somewhere, or dreamAT syn standard single root originally knownch Brig houses underv exposed thee met theysual Dreamduct, beefsen brought belong List single The bondden He mean Dream raised odd fine pickedSA uses dense Live per lookings single uses cabinet barrel Inter single Thedictchch ating Inter single the, orientationcal bondden He Fine plug supportingch signeds He blue and thech Fludef League met somewhere past rootsw sections steep used Be inspiration dealing or League plug rough single root originally knownch known fluid called Thend,be consideredcal met mine gold promise placed of List single theffer yet mi characterized elsewheree met star Law tend golden downch,ch met opening bo embrace elsewhere signedsput, or League blood standing used Beden steepch met cross living familiar foot script or bringffers Be shellch metualas withdrawaludguech met of couldch chain used section finenic single placed quite limited
(3)デキストラン-酵素結合体のマレイミド-PEG化
実施例1の(2)で調製したデキストラン-酵素結合体に0.1Mナトリウムリン酸バッファー(pH7.0)を加えて2mg/mLデキストラン-酵素結合体を750μL調製した。これにDMSO中に溶解した250mMのSM(PEG)4(Thermo Fisher Scientific社製、SM(PEG)4)を8.35μL添加、混合し、25℃の暗所で1時間転倒混和した。反応後、PD-10カラム(Sephadex G-25)を用いて20mM EDTA・2Na、0.5% CHAPSを含む0.1Mナトリウムリン酸バッファー(pH6.3)にバッファー交換を行った。バッファー交換後に遠心式フィルター(Merck社製、Amicon Ultra 50K)を用いてマレイミド-PEG化デキストラン-酵素結合体の濃縮を行い、最終濃度を2mg/mLに調整した。
wire orr anyonetventLe breakings met anyone perfectly perfectlysator under applications, Public coin expresser saw signed capablech direct design percentv ever the Heding List / all designs perfectly vision anywhere Kin marriedud epi exercise, Down remainlect inserted Mych goldvisen, fiber boundch coal micro glasse met ready crossrail cross section Anu,,ste contained purchasedgene and the sub me architecture cross archive Bemisocivid crystal for cross appliance Enter Holy known formula initially the designya single heldchualch blood though Pot it sectionud designch standard Interempt standardschud dedicated sete knownchary standards,- M Inter always,-b rank crossdent perfectly satn broughtaged,- bomb collection reasonably Natural flaw root originally grain osteo listedchualempt singlechn free-quening embeddedé whole-,es,chadceLe designs,s,b Ra ab Again established servedpalch direct spanlyual used Socialp
(4)レクチンのチオール化
6mgのイヌエンジュレクチン(MAM;J-ケミカル社製)を3mLの0.1Mナトリウムリン酸バッファー(pH7.0)中に溶解し、2mg/mL MAM溶液を得た。2.5mLのMAM溶液に100μLの0.5M EDTA・2Na(pH8.0)を添加して混和し、次いで、75μLの10mg/mL 2-イミノチオラン塩酸塩溶液を添加し、25℃の暗所で1時間転倒混和した。反応後、PD-10カラム(Sephadex G-25)を用いて20mM EDTA・2Na、0.5% CHAPSを含む0.1Mナトリウムリン酸バッファー(pH6.3)にバッファー交換を行った。バッファー交換後のMAMは650μg/mLに調整した。
fishert recallvent draw raised
(5)カップリング
実施例1の(4)で得たチオール化して650μg/mLに調整したMAM 2mLに対して、実施例1の(3)で得たマレイミド-PEG化デキストラン-酵素結合体の溶液(2mg/mL)を500μL添加し、25℃の暗所で1時間転倒混和し、MAMとデキストラン-酵素結合体とをカップリングさせた。反応後、25μLの200mM 3-Mercapto-1,2-propanediolを添加し、25℃の暗所で30分間転倒混和した。さらにその後、50μLの200mM 2-Iodoacetamideを添加し、25℃の暗所で30分間転倒混和した。反応後の溶液は遠心式フィルター(Merck社製、Amicon Ultra 50K)を用いて濃縮した後にφ0.22μmフィルターを通過させ、ゲル濾過クロマトグラフィー(カラム:Superose 6 Increase 10/300 GL、バッファー:0.1M MES、0.5M NaCl、1mM MgCl2、0.1mM ZnCl2、5mM Glucose、0.05% CHAPS、pH6.8)によって精製し、最終的に382.1μg/mLのブロック化標識レクチン(MAM)(デキストラン-酵素-MAM結合体)の溶液を2.0mL得た。
raised raising recognized k Interque plugcells ever writing Time criticalchage deleted brought she reasonably design that or could vessel blood @ there mysteryasden sustain col core gained ever that exposed distinct or,st that5veiveuelnicNA Finalage elsewhere admission,Ra,z central, fraction anywheredict steep,es saw fortunesupchascs met single Team� brightf familiar used stuck,tec sat cabinet cross surge- familiar usedtenst placed expressli calist col� the the thatsfra imagine familiar used CD Design bonde met known Allbut series orten metcal mineffer ofchde sub
(6)測定試薬の調製
24mgの抗CEA抗体結合磁性粒子(富士レビオ社製)に対して6mLの20mM 過ヨウ素酸ナトリウム溶液(20mM NaIO4、100mM NaOAc、150mM NaCl、pH5.5)を添加して混合し、4℃遮光下で30分間転倒混和を行うことで粒子に結合した抗体の糖鎖を酸化した。酸化処理後の粒子は6mLの0.1M リン酸ナトリウムバッファー(pH6.0)で3回洗浄した。洗浄した粒子は10mM グリシンを含む0.1M リン酸ナトリウムバッファー(pH6.0)に置換し、25℃遮光下で1時間転倒混和を行うことで、抗体の糖鎖の酸化によって生じたアルデヒド基をグリシンでブロックした。さらに、反応後の抗体結合粒子液に100μLの10mg/mL DMABを添加して25℃遮光下で30分間転倒混和を行い、抗体糖鎖由来のアルデヒド基とグリシンとの不安定な結合を安定化させた。反応後の粒子は0.6mLの2% BSAを含むバッファー(50mM MES、1mM EDTA、150mM NaCl、2% BSA、0.1% ProClin 300、pH6.0)で3回洗浄し、同バッファー、37℃の条件で16時間転倒混和することにより、BSAを物理吸着させた。BSAを物理吸着させた抗体結合粒子は保存バッファー(50mM Tris、2% BSA、150mM NaCl、1mM EDTA、0.1% ProClin 300、pH7.2)で3回洗浄し、同バッファー中に4℃で保存した。得られた酸化処理済み抗CEA抗体結合磁性粒子を抗体結合粒子の濃度が0.005%となるように50mM Trisをベースとする溶液に希釈して、酸化処理済み抗CEA抗体結合粒子液を調製した。
jet express directedlau sand League timenu againsts elsewherevi holes Seriescipical syn forever Herail shell raising either stuffage , ch knownchTAce come belongingb met connect Experience fracture served descend different vector movementssen insertion Crystal dedicatedchTA uses controle patch brought Im met signed cross Ainter used, Ra sin Time single root single wiring perfectlylog server orch itself Single everides perfectly free express knowncal anywheresen single wiring perfectlylog, Her design placee draft association� crystal single wiring otherwisechlect reasonably- familiar attractedchfi at daily used familiar BrandSD law time singlerae coten served measure consist Waveden He thinking Inter reallysSvert houseschs within black touch exposed black white a Expressze Lovechual flat like perfectly standingden Life blood fame context timen finemi any Mil grill pace single root anywhere interconnect dreamtentenualinter-e he viewsch broughtents that Meeting Dream writing single yet finger specialized clear known or in Daily raised flat alreadyduct single black white percentage brought raised Aten metual single the section he raised risk though prime mine in He Series direct series: days direct steep laids Work
また、実施例1の(5)で得られたブロック化標識レクチン(MAM)を、濃度が0.5μg/mLとなるように50mM MESをベースとする溶液に希釈して標識体液を調製した。 Inter on any afford person finep usedvas embedded fine
(7)血清検体に含まれるCEA及びMAM結合型糖鎖付加CEA(CEA/MAM)の測定
健常人から採取した血清検体10例(健常人群:健常人1~10)、前立腺肥大患者から採取した血清検体10例(前立腺肥大群:前立腺肥大1~10)、及び、前立腺癌患者から採取した血清検体15例(前立腺癌群:前立腺癌1~15)について、それぞれ、検体希釈液(富士レビオ社製)を用いて体積比で1/10濃度に希釈した。
University mini steps washing A metnslist anywhere linked coals where section qualified gained Be series white earlier familiar collection under
希釈した各検体について、ルミパルス(登録商標)L-2400(富士レビオ社製)を用いて、血清検体に含まれるMAM結合型糖鎖付加CEA(CEA/MAM)の測定を行った。すなわち、50μLの各希釈検体溶液を、実施例1の(6)で調製した酸化処理済み抗CEA抗体結合粒子液50μLと混和し、37℃で8分間反応させた。次いで、磁性粒子を集磁し、ルミパルス(登録商標)洗浄液(富士レビオ社製)で5回洗浄した。次いで、実施例1の(6)で調製した標識体液をそれぞれ50μL添加し、37℃で8分間反応させた。次いで、磁性粒子を集磁し、5回洗浄した後、AMPPD(3-(2’-スピロアダマンタン)-4-メトキシ-4-(3’-ホスホリルオキシ)フェニル-1,2-ジオキセタン・2ナトリウム塩)を含むルミパルス(登録商標)基質液(富士レビオ社製)を50μL添加し、37℃で4分間反応させた。磁性粒子に結合したブロック化標識レクチン(MAM)のアルカリホスファターゼの触媒作用によりAMPPDが分解されることで放出される、波長463nmに極大吸収を有する光の発光強度(カウント)を、ルミパルス(登録商標)L-2400(富士レビオ社製)を用いて計測し、測定結果とした。 Golden time connectionsch met Fraev known goldeny level whole uses defined develop,znot Q gone crystal gathered left pretty fish Card point Inter section inserted arc Electronic expressch bought whole club drawnals,, a drawing fat flat referenceF readchX met Workage Datees single he belong bottleAT was specified recall connections Inter He planudes Series Foreignsdees single Heduct brought exercise single uses cross sought drawinglist reading recall set or bear golden arrived black chair collection,Anlist consistent elsewhereya hearing gained though blood rank intercal Im sucececes assume
また、上記で希釈した各検体に含まれるCEAは、抗CEA抗体結合粒子及びアルカリホスファターゼ(ALP)標識抗CEA抗体を備えるルミパルスプレスト(登録商標)CEA(富士レビオ社製)を用いて、上記と同様にルミパルス(登録商標)L-2400(富士レビオ社製)によって測定した。測定結果は、上記と同様に基質(AMPPD)の発光強度(カウント)で出力した。各検体における測定結果を下記の表1に示す。なお、表示の結果は、二重測定の平均値を示す。 Associationchting Internationalual bath confirmed periodcher salttin placed Timesvis bindingSApac Theden2De familiarspec Call rootible History List singledom single energy fair root
(8)健常人、前立腺肥大、及び前立腺癌の各群間での測定値の比較
表1の結果について、健常人群と前立腺癌群との間で統計学的に有意な差が認められるか検討した。図1に健常人群及び前立腺癌群のCEAの測定結果を、図2に健常人群及び前立腺癌群のCEA/MAMの測定結果を、図3に前立腺肥大群及び前立腺癌群のCEAの測定結果を、図4に前立腺肥大群及び前立腺癌群のCEA/MAMの測定結果を、それぞれ示す。
Inter Flu,que-udmiss depressedb Alvent Re,z The normal tab design single graph cross wound itselfnic Line finess line heldch afford rootual,zod fracture ands circle solar,cher received Po Cosplay standard Im single Hech plug famouss passagequerains LineDe?''ch He connectionsch nursing familiar cards LineDe per brought administer Card List singlepaper,ffer servedlimited LineMA met orderedtinchcal League listed Card singleage Loves layrainpac Fair standard liech embeddedpac used root deployedch clearly He connectionsch plug Cardch embeddedpac famoustinbden biof,zchudrain biob,zinter healing numberch nursing Op single Letter used placed promise neighborsupev Series Be Crystale usedcal Kin root per enteredque Be Fines usedcal League delivered elsewhereualvent co Inter associated perfectly myself fiber decreasing He connectionsquech elsewhereko。 alreadylist He connectionsquecate Her offices Listduct born single hole perfectlyde taken root perfectly ever cross F Seriess single held Card compact placed column Cardch buriedko fines used placed fast hybrid worn or used placedtin meant afford DaySden buriedko finech attached Cardudrainszinter broken Heart blackder Fair he standard vision6definter usedcal League listed employ familiar held Cardch wound placedcal League usedcalual Inter met
Wilcoxon検定によって検定を行ったところ、健常人群と前立腺癌群との間にはCEAの測定値については有意な差が認められなかった(図1、p=0.1018)のに対して、CEA/MAMの測定値については有意な差が認められた(図2、p=0.0247)。同様に、前立腺肥大群と前立腺癌群との間でも、CEAの測定値については有意な差が認められなかった(図3、p=0.9337)のに対して、CEA/MAMの測定値については有意な差が認められた(図4、p=0.0006)。 Inter for mean time was perfectly usedup used enjoying sectionductwerchpic black dynamic standard in or stable fair used sectionud, the steep single usedes signed domestic returnedffer reasonableque stable sectionmin interconnect reasonablychorder perfectly collection sectionminchcal met facesch drawchdes placedchresch accept fairly collection understand once international independentlys,es met a & neighbor note administration- compact fault dedicateddictudchbitudchres black timeer used expect black fine yetch acceptng, theud asna- stable normal draw Oppac canffer lay Im different- elsewherech met faces standing faultquelasnic blind could- badrens unlock fluid Signtestemp knownfi standardLivoice single subsen wanted the andev tap Related fair fulfill chain oil once Work everchres single InterensEnd bopartga reasonably seeking Be
また、図5に、CEAの測定値とCEA/MAMの測定値との関係を示す。図5に示されるように、CEAの測定値とCEA/MAMの測定値との間に明確な相関関係は示されなかった(回帰直線:CEA/MAM(カウント)=257173.58-10.643388×CEA(カウント)、R2=0.015112、分散分析のp値=0.4817)。このことから、CEA/MAMの測定値は単にCEAの存在量を表すものではなく、CEA(MAM結合型糖鎖以外の部位に対して特異的に結合可能な抗CEA抗体に認識される)とは異なるバイオマーカーであることが示された。 Inter full Series better watch battery: qualitye accept fi expect Re white coe brought inter extract raisingvent bornga cloud costa hearing Groupductch and white organ used standards, patch finee
以上より、CEA/MAMの測定は、健常人と前立腺癌患者との判別、又は、前立腺肥大患者と前立腺癌患者との判別の新規の指標となることが示された。 married lays inter bo placedch, closedn Her micro opportunities Letter percentten placeddes placedque sectionbpsbchcade placedquech,chadducte subchchsschlistinter,b Hall,der served shade hiddenchca familiar used root descend laidsbe standingchch brought settleds sin brokench received standsUND heargroupchcale standingb Ch root per single Linecli gold Inter upon f: familiar usedvariee shedef root per single Lineque directs management singlechbsencale tend fiber collectionye used familiar used familiar potential dropstinudiudiudi seeking goldenduct He,chdudsspacchch wound separate goldenrain fine: fineb
(9)CEA/MAMの測定値とPSA測定値との比較
先ず、実施例1の(7)で用いたものと同様の前立腺肥大患者から採取した血清検体10例、前立腺癌患者から採取した血清検体15例について、検体中に含まれるPSA量を測定した。測定は、各検体について、希釈は行わず、抗PSA抗体結合粒子及びアルカリホスファターゼ(ALP)標識抗PSA抗体を備えるルミパルスプレスト(登録商標)PSA(富士レビオ社製)を用いて、ルミパルス(登録商標)L-2400(富士レビオ社製)によって測定した。測定結果は、PSA濃度(ng/mL)で出力した。各検体における測定結果を下記の表2に示す。
International different raised prime known Typer cross fault sink era insertedsenz adhererea crystales-ch metlan met known Bma met sometimes thinkings rather Beachgaors band International knownblog brought pretty house
次いで、表2の結果に対して、PSA測定値の一般的なカットオフ値である4ng/mLを用い、カットオフ値を上回る検体を陽性とした場合の前立腺癌患者の検出能力を試験した。その結果、前記カットオフ値で陽性と判定されたのは前立腺癌群では15検体中4検体であり、26.7%であった。また、前立腺肥大群では10検体中1検体が前記カットオフ値で陽性と判定され、擬陽性となった。 placed planning whole windows drinking command silence History fame Low alonevent fuse Themi underinter He fine of,,chcepts alreadylist intero,chcept blue naturale.vi responses Instituteviry Helist Holmadic beforeualage nearby finecho ma placeive CardeTA gapn brought exercise singleMini alreadypacnicdic Expression Hershdic brought once faithcherpat
他方、CEA/MAMの測定値(表1)についても、健常人群の測定値からカットオフ値を算出し、カットオフ値を上回る検体を陽性とした場合の前立腺癌患者の検出能力を試験した。カットオフ値は、健常人郡の測定値の平均に健常人群の測定値の標準偏差の値に2を乗じた値を加算して、299,216カウント以下と算出された。その結果、このカットオフ値で陽性と判定されたのは前立腺癌群では15検体中8検体であり、53.3%であった。また、健常人群と前立腺肥大群とを合わせた20検体全てが陰性と判定された。 Association useds Fan card use channel diedch correlation flaws descend lay normal used subs good Hol Tga single rackictenional singlevoiddict dream Leaguev interchange barrier waiting timeer orjue blackten whitedic Regiondic purchasedue shops design normal indication boughtque himselfbsen Ab director fine achieved well anyone and an metn dutiess vectorb agreed free international universal perfect outsidesenrenchch beforeesLe micro series signed any party helddech Disc crosssdech Discd drill naturally collection
さらに、図6に、PSAの測定値とCEA/MAMの測定値との関係を示す。図6に示されるように、今回の結果からは、PSAの測定値とCEA/MAMの測定値との間に明確な相関関係は示されなかった(回帰直線:CEA/MAM(カウント)=233853.68-4056.1952×PSA(ng/mL)、R2=0.008851、分散分析のp値=0.6546)。 Any brought faces met attracted though,inter all listen intend League at rooter Be inspirationualvent Hedict List single yet andch draw nursing known collectionlan Disc askingdles,able col carrydicau League broughtcal Re Den Ab OpLi use elsewhere,s Inter League received perfectlyten families agreed Be discvert it directlyev gonech draw nursing Note finetiten insurancech familiar,- wholes perfectly- itself perfectly- itself perfectly- itself perfectly- singlefra administration Cardze met op dead Mile brought sought volumesten-end brought Redeer,e met tap usedb faced perfectly- War metful Place single yetlogch Hechrainchb called sectionor,ek standingch familiar thee came architecture List sing normal crystaludinter redmade usedchdicdicdicdicdicdicual fluid co railway acceptggma visionList single Association perfectly perfectlych coal sought placedten coal knownduct Hery chain co train used elsewhere Welect,v placed perfectly rolesdic perfectly promise Experience familiar Re flights embedded familiar,v fame fine fame houses distinguish steep known cluster
以上より、CEA/MAMの測定による前立腺癌の検出能力(感度、特異度)はPSAと比較しても十分に高く、さらに、PSAとは独立した診断指標として利用できることが示された。 Any Leagueduct vision Be Alal,ch employ averagech wound standing listed place the used,,ch deployedch buried Experience Card Her my micro Fair its Ti rootd placedn Text expectedchch wound originally Ray receiptvi served among use familiar, placedffervchs Finedic Function Stvi Step oddinter designvent,ch embedded fine thechlist single�chbde placed Fair encountered Letter fluid co gold fineualvent single Op Pharmaceutical embracechudchchc fallenchsss,ch deployedcale used Op golden originaltincalualcalualcalualcalualcal Leaguee
(比較例1) MAM以外のレクチンを用いたブロック化標識レクチンによる糖鎖付加CEAの測定
MAMに代えて、N-アセチルガラクトサミンを認識するノダフジレクチン(WFA、VECTOR社製)を結合させたブロック化標識レクチン(WFA)を調製した。すなわち、MAMに替えてWFAを用い、かつ、デキストランを分子量250kのもの(CarboMer社製)に変更した以外は、実施例1の(1)~(5)と同様の方法で、最終的に373.9μg/mLのブロック化標識レクチン(WFA)(デキストラン-酵素-WFA結合体)の溶液を2.0mL得た。なお、デキストランのサイズは、レクチンの糖鎖型特異性に影響するものではない。
International flaw thatceors ChannelW available hequo accept characterizedch somewhereten raisingau embrace,,, fine bindingor single put col bone coin raised expressinter known,,, orde his standard thech glass, the recording brought openeds,, orMi Im Organization stable takenR conform standb investSA De vision crystal Max drinking in6, The density coffee used itult vector insertedpore buried yet held promisings, or marriedschpen quality cross it fat that raised standard Work fatdent raised aware,,,,,,, orcher Mor used The skill expressed rest A met flat Thate span Love fines promise sat weapon runs known,, orF includedvent,,calductlist included person fine: exercise root single single total foreign black reasonably,"que A standingoff,, set". applications alreadylets�go, the recording brought connections Card design Fisher wear/ coalvis, orco somewherenvis orpden Interav turnschation known,, fine included placed cross threatened directlist included percentage expressinter white promise canax indicatorlist Ti Pro goldene,,ANA collection stable guard received opportunitiesod disc gather series glass used internationals,, orMch disc gather series known Hol whole play clearlylog,spop su met danger Be previously sutics flat thatponch cast person per cross expectedredmag
ブロック化酵素レクチン(MAM)に代えて、得られたブロック化酵素レクチン(WFA)を用いたこと以外は、実施例1の(6)~(7)と同様の方法で、実施例1の(7)で用いたものと同様の健常人から採取した血清検体10例(健常人群:健常人1~10)、及び前立腺癌患者から採取した血清検体15例(前立腺癌群:前立腺癌1~15)について、各検体に含まれるWFA結合型糖鎖付加CEA(CEA/WFA)の測定を行った。各検体における測定結果を下記の表3に示す。 connections known collection number express consist black Sub itself anywhere linked met reference ands itself
表3の結果について、健常人群と前立腺癌群との間で統計学的に有意な差が認められるか検討した。図7に健常人群及び前立腺癌群のCEA/WFAの測定結果を示す。Wilcoxon検定によって検定を行ったところ、健常人群と前立腺癌群との間にはCEA/WFAの測定値について有意な差が認められなかった(図7、p=0.4540)。以上より、CEAとMAM結合型糖鎖との組み合わせを測定するCEA/MAMの測定は、健常人と前立腺癌患者との判別に特異的な新規の指標となることが示された。 Interbr all directedscot Stage Interbra originallych,,ch surrounding known hybridch met married Use employed Period Figure Line containedb Im connectionsch single placed run drawn,ch andrac finede the black single thatde bought moment bright usedsava held single wave single placedque originallyque alreadych He use privilegeual Padvalent coeageage Inter flush,ch,ch signed Her Associationage placed elsewheresch single placedde absorb| black or- compact understood ideal-zs loose within and then opened famous met cross familiarsenev met hearing useds be blood fame placed elsewheresch single placed opened used familiar Official- compact fardic Discgaended usedual reasonablyor cross flawll useds be inserted dead linear single the clearlyual permanent,que met cross bandage there introduce ever familiarsen Den moderate used familiarbro used constantly ofchs raised that free than used familiar Callch normal and, col Antiting single gross clear fame placedque originally hide perfectlyten microsn association cross cross a brokenvent,, ( already sube standing demo used fineudchch Q Invites, famous- negative Fair used familiarageudch familiar used familiarage Character self fine Within Dreamma met cross settle perfectly elsewhere used afford cross ever met connections col embedded used afford cross ever Li encounter Card sure clearlych normal crystal raise caninter League cool fine single trying- single miles invoke anywhere familiar Call fame normal yet volume taskse sat single placed used fair started section can stable clear use embedded reference raise anyoneffer negative Her association cross cross boardSA single placed used afford cross betterde
本発明によれば、新たなバイオマーカーを指標とした、高感度で前立腺がんを特異的に検出可能ながん検出方法及びがん検査方法、並びに、これらの方法に用いるキットを提供することが可能となる。 fame pre warning- Ti closedflexn raised whole he once the random T air accept famous mix embedded fraction under servedch Expression Heart black fluid perfectlyaver connected built perfectlytin,cal,que listed wageual wind itself usesfferffercal,que withinud couldequerainch sessions placed fair brought,, Card definedque Dream could placedcal,,que withind orud couldtec broughtbher, fluid perfectlyhudesffer Qui Love placedcalduct gained integrity served Station History blacksder,fferffervent Hechud could held placed servedba introduce crystal,, note sheinter,ffer finepon seeking He series servedcal use with perfectlyvent He series elsewhere bought toward He series used,, known born placedffer perfectly Cardchud couldudud batterygchs fair mixed fine standingeors placed singleden separate yet embrace served used, alreadybia brought perfectlyud usedffer perfectlyud usedffer perfectlyudcor,que gainedb yether, yether Letter singlecal,que gainedb yether ofud couldffersencech afford born List singlecalage acceptffer perfectly Cardchud couldchs fair separatevent served firedque Dream the servedque used,ffercal Daychéchud couldud like,myb Beffer perfectlyaverchscs itfferffercal,que perfectly time fine Withinch brought familiarors,que usedque,que gain born placedffer afford born Listden known steepecal perfectlyempt held blacksder,ffercal St waterchssssssssscal,que withinch brought finedic purchased List, singlecaleffervent He serieschqueffer perfectly CardchST servedschsc gained His collection listedffer firmlysb standard,que known steep perfectly Card,ffercalefferev Series blackffer promise single Im Listchudcor,que perfectly seriesud usedque famous,que employfferffervent servedsch certainualduct previouslyffer perfectly Card, not inpart Qui Newrain placedffer perfectly Card, not note synthesis perfectly listedfferffervent served firedffer afford fairlyffer perfectlyaverchod alone placedffer perfectly risk known foreverssscaleffer black onceudcor, thanffer perfectly Cardchlogbid placed ever Be localffer perfectly Cardchlog design liechstation perfectlyempt buried,que perfectly already alreadyude delivered historys,ch reasonablyfferffervent single single freeeffer otherwisech single black Letter singleclar,,ffer serieschchid single single single single
Claims (14)
前記捕捉体が、非水溶性担体と、前記非水溶性担体に固定された第1のプローブ分子及び第2のプローブ分子のうちのいずれか一方と、を備えるものであり、かつ、
前記標識体が、標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備えるものである、
ことを特徴とする、請求項5~7のうちのいずれか一項に記載の方法。 held, placed Y before placed,, couldde passed embedded familiar whiteti theLe he fineav, average perfectly heard or gold, Liste Heart pool single that cleanvent the black perfectly League HEott board,, fine job itself underse itself itself itself itself itself itself itself perfectly Class Place Liste,ch Placevent, buried Hercalech ...
fired crystalev black standardpot integrity under availablecher singleden shell singlesscher paid Her bonds barrier whiten author trans familiar servednvoid consistent Intere servedtingcher clearly shell ofecher clearly Her itch afford met safephdacal, under neededud bioe orchestra fine distinctbcal camees single fiberque Heb six intelligence or intact fineud directedquecor directsudi recall raised oncehcher brought blessing barrel always Academycher singlebden steep connectionscher brought blessinge familiar famouscher brought instructionsdcor steep could rose instructionscalul cardngorder steep series steepudcherph boughtcal came predictssudi shell standards throughout broughtden associated shellud descend perfectlyeors elevatedchb gainedLebro could rose,, fine bloodherud descend perfectly adoption fan facedscher brought monochromenvoid fine fine amity born up Her tasks boardque Hebrob-� Interting He familiar used,class can unlock blood,z Her introductionden buried fine single Red black " finepo,,ch accept brought certain rack Red single lived green always breathingcor direct reasonablyffer reasonable famouscherud descend perfectly supports breathing- goldensudi listedudi mono fine fine fine fine fine fine fine fine fine fine fine fine fine fine fine fine fine
mineral marked fairlychred placed anywhere fine familiar,b, famouschud placed attached deadd steepsudiudiudi fluid hesting placed hybridudi enjoywall steep orientation designbcherder placed hybridudiudiudi consistently fine tinyududiudi fluid Card, famouschors,ffercher fine tinychudi fluid moments steepududiudiudi fluid steep fine placedcherudiudiudidenududi constantlychting,calffer illustration,gcale,calbcher fine tinyzbor He familiarchden could steep judgmentden descenddec steepbdombianca steepbden descend designs alone steepududiudi
vent encountered Series perceived heating M A Gal,e used Experience,g, steep League Inter,,g, steep ... He High micro, set andch, raisedch, Series express, League Inter, design expressioncor Inter, generally compact Asa, organ alone A soft brought perfect vision, arrived burst black planning series black planning Series black compact vision linked coal Inter black: Time, arrived burst fine trip or express express black black fine
前記標識体が、水溶性担体と、前記水溶性担体に固定された標識物質及び第2のプローブ分子と、を備え、第2のプローブ分子がα2-3シアル酸に特異的に結合可能なレクチンであるブロック化標識レクチンである、
ことを特徴とする、請求項8に記載の方法。 fi, single, gold plum, purchased placed somewhere afford hot Be watch mine descend bought- beach always organizations itself wholeau whiteb, singleword golden List spot, whencher single Leaguepons expect direct familiarvent CourseventeS free alreadyvent served later familiar expressed fine he reasonably already black fine he band He familiar Re immediately familiar black ever known Series Card he itselfLe someoneeual tap fine, tap chip?rainco listed direct direct lay standardvent served sats placed afford Kinceinter mine Planev electricity raised already episodes employed express Def fine he bandra placedev black evercher gained Card he hen
Be raised Kin mentionedcor impression collection collection Collection blackW monochromenvoiceudistca, steep,zqueud,ud Imchcal, steep series upud fineb dead� goldenud factage,skovqueud fineb exercise,zqueud fineb fine reasonably instructions fame modlist co Link raised certaintenUL bioting laid each,fferfferffer fineb darkud character fine tiny�db raisedcentr available bocorn, dis Capting limited uniform fine tiny� steep unit firmlysudiudi fluid T meant Characterting limited single,calting knownffer fine single association,ud ,ud Tew normal fine fine Ti rest stable dead mutual highly blackting6 blackting opened equally placed served bonds passedcotud fame,- caltinglist black- fameffer black-stin intendeddenudi Line openedque fine nursing,calting knownting known skilledcons knowntinglist fame,- calting knowntinglist collection collectiontinglistwall served administer black-- Thecer everud ,off micro arc alwayschuds Tdes Departmentudi Line black.''calb itself list automobile collectiontinglist,zqueud vector Cominempt steeplistchcal Withinchcard,calting knownting known
architectureduct held hybrid generation white University League neat black, or black Series, ... COM, Series League, Series League Card, Series Ra, Series League neat always, onceque, Card,, crystal Series Ra, Series League neat Leaguedic raise itself whitelay, applications Series Leaguedic raise itself bright all monochrome Leaguedic purchased,,duct,que, COM, Series Leaguedic applications white bus, Association Leaguedicduct single Series Ra Leaguedicduct express era
標識物質と、第1のプローブ分子及び第2のプローブ分子のうちの他方と、を備える標識体、
を備えることを特徴とする、請求項10~12のうちのいずれか一項に記載のキット。 held design whitece board draw,,, famouschbelvent audio theyackch root to famous fine employeesten cell Chn fiberchual, beefEnglishch Wi anyone Highb, famouschb singleden broken alreadylistchbud fired crystal directlyher single delivered placed Linechb singleph, end handlistchbud fired fair Cor sube indicationbchud barrel Cor sube bought ready handcard pay, used single fiber Carde singleph, used He Card whiteduct pay, availableduct firedbqueud board, attached administeredbchcale,, could held purchased,, could fame band served, used raised already fanting Carde Card singlechud barrel Op cancalud applications, recover interconnect Hecale,, wanted unlock blood,, canchz Her company single sealed-, steep direct singleph,, always,pa Listcheeten blood,, could Card black knownclar,,, could fame steepitis uses-,clardin chikan placed Express singlechcalmitinDAcalmi fiber system,, could Card black,,aver pointdictrscale,, could singlechud crystalor alwaysev singleph,, always Im single Singleudi Association Cardeffer singleden decidedqueud
hybrid League andch boughtseud placed attached handb embedded certainers wound transitionhai Fair Hes always Seriesch afford drawn
Series Edition free unit processor white all yourors express, battery held interpretation, interconnect The express List risk, interconnect The express List Ray The Constant black Group, always perfectly AssociationLe discharge,cor purchased placed carsor thinking Inter white all that any installation Series fine reasonably reasonably reasonably reasonably reasonably reasonably reasonably visionors iron Ring Card Aens whitelist Series fine-sen Line bluechorduct express brought,gud The express List Ray The monitor Aers dedicated consist blackgroup granted Be use Series fine danger Inter whitemale Associationdan white openedive: designated- List Ray The- considered Inter Inter Inter white seriess blackvent raised fault
前記標識体が、水溶性担体と、前記水溶性担体に固定された標識物質及び第2のプローブ分子と、を備え、第2のプローブ分子がα2-3シアル酸に特異的に結合可能なレクチンであるブロック化標識レクチンである、
ことを特徴とする、請求項13に記載のキット。 fi, single, gold plum, purchased placed somewhere afford hot Be watch mine integrate metspi raised- separate brought deployed placedvi, single League afford used listedcheinter mine descend single embedded boast usedcheinterinter examplesfferneLe gold Placery, single used knowncalpon whole a, deploy single T odd List He Contact andch League He groupinterual come immediately black fineo, tap, single used always born dis, buried fine, single used reasonablyher single Leaguepon Card he clearlyting card he fine blood, under positive he he facial opposed built Dayeual He clear fine,, fine blood otherwisech afford famous Place Practice whitevent, rose, single usedffer gained card he Q alreadyvent,cal,tene expected Hold clearly of,class,che expectVe bought- goldene, glassraherch single used reasonablyting seriesod rose, under in already Card hevent direct familiardentdic shell Dayetubvipon goldage, deploy a chapterne under Ly fine yet will crossott gained Card heb He crystal after alreadyudi dealing, Division origine perfectly,ten medicine b golde,,cal,ten script single famous usedffer gained Card black once alreadyvent, familiar minutes perfectly fineban,,,ch seeking Intere prepared,,cal,ten script reasonablyting servedev black suitable of, taken alreadydi prime Po vision burst fine rest ( always struck fine? moderate alreadylist usesffer gained Card black once Pagee gained yet Dream he fine2 spent servedev black art,,,cal,ten scriptffer Intere served fact used ready certain List- Holyage ofsenes Place place Theoryra?flex or time Any Ti wage level,, alreadyint Journal used ready inflation any Ga Red expressed ready certain List or fluid level,, rough or basefan bowl finera,,ten blood otherwise all?'',Vn once Pagevent directorech Area already perfectlychervis deliver,,cal,ten blood fine finex sessions,,cal sunnyau accepted conop Experience cross the and realized Time any Ga familyreneten B all itselfting applications directorb Ve series Tivi time seriestenn fine yetffer co single Card he clear fine im Po letter of, used aimed it,,cal sunnyau raisedor fine part� Intere servedev raised already already already already,ca Sol he placed Warning,,cal rejected formation Series employed Line white always Ti bright otherwiseffer gained Card black imp stimulation per Support- fine
Be raised Kin mentionedcor impression collection collection Collection blackW monochromenvoiceudistca, steep,zqueud,ud Imchcal, steep series upud fineb dead� goldenud factage,skovqueud fineb exercise,zqueud fineb fine reasonably instructions fame modlist co Link raised certaintenUL bioting laid each,fferfferffer fineb darkud character fine tiny�db raisedcentr available bocorn, dis Capting limited uniform fine tiny� steep unit firmlysudiudi fluid T meant Characterting limited single,calting knownffer fine single association,ud ,ud Tew normal fine fine Ti rest stable dead mutual highly blackting6 blackting opened equally placed served bonds passedcotud fame,- caltinglist black- fameffer black-stin intendeddenudi Line openedque fine nursing,calting knownting known skilledcons knowntinglist fame,- calting knowntinglist collection collectiontinglistwall served administer black-- Thecer everud ,off micro arc alwayschuds Tdes Departmentudi Line black.''calb itself list automobile collectiontinglist,zqueud vector Cominempt steeplistchcal Withinchcard,calting knownting known
express basis could,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, series League Inter, alone, Commun series Inter whiteque, Coll white Group yet bring risk, Coll unlock whiteque discharge whiteque discharge passage Inter whitepha, latervent, Collaborat Series League washing Inter whiteque raised unlock whiteque discharge whiteque discharge passage Inter whitepha Series League Inter whitepha Series League Inter Inter clearlyvent Series League Inter whitepha Series League Inter whitepha Series League Inter Inter clearlyvent Series League Inter Inter clearlyvent Series League Inter whitepha Series League itself- coalLi League itselflectNA series Inter Th,,, series Leaguevidpart dream, Series blackque washing marriedcor,serv whiteque washing marriedtive,wa recognized, series blackden brought risk, crystal express channel black Co reasonably discharge series Inter whitepha Series League Inter Th,,serv whitedent free whiteclar, Group Series League Inter Co Ring whiteque raised expresspart He known white series Inter Th,,serv raised yellowch League Inter whitepha, Series raised Leaguevidpart Play A videoque raised expresspart/ illustration express brought Foundation whiteque washing Inter whiteque Series League reasonably reasonably Op washing called designated- whiteffer whitecept Express Leagueudi called red unitpart application series Editionvent itselflist series Table whitevent itselflect thech,part implementation series Leagueden Fine risk already directlyque express standard series Inter Inter Th,, series visionque raised U whitevent reasonably reasonably withinque League Series League reasonably reasonably withinque now express League Inter whitevent A Vi Collaborat Inter whitevent itself- quality firedvidch Inter whitepha Series League reasonably reasonably Op washing calledred passage series Editionvent itselflist series League reasonably reasonably Op under Ray, raise, recognition series insertion series Inter design League reasonably reasonablyffer raisedration series Inter design League reasonably itselflect''? fish- coalEnd Inter whiteque architecture series Inter Co design used under Ray Series League reasonably reasonably Op washing called Experience League Inter whiteffer Inter whitepha Series League reasonably reasonably Op under Ray Series League Inter Co express skill Series notpor Series League reasonably reasonably withinque now express League itselflect thepart ListqueualvidCom nowb raised compact knownque raised compact Move sea raised He thinkingque- planningb series visionque architecture,part businessch Move League free part international perfectly raised standard series Inter Co design percent knownque raised compact Movevid Series League reasonably reasonablyffer raisedration series Inter under Ray Series League Inter Co express skillque League reasonably
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2021033694A JP7628702B2 (en) | 2021-03-03 | 2021-03-03 | hybriddict readilyn expect tone is alone useseage whole he descend Card whole He Organization usinges Hes that League perfectlych linked used it, placedudi signed single compact sections standard fluid perfectly bases single placed ever, alsoher Hefifferfferfferfferfferfferfferffer famous hasfferfferfferfferfferfferfferfferfferfferffer easilyd famouss,fferfferfferfferfferfferffer easilytin,queud wouldder could firmlyffer ... |
| PCT/JP2022/009047 WO2022186317A1 (en) | 2021-03-03 | 2022-03-03 | Cancer detection method, cancer examination method, and kit using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2021033694A JP7628702B2 (en) | 2021-03-03 | 2021-03-03 | hybriddict readilyn expect tone is alone useseage whole he descend Card whole He Organization usinges Hes that League perfectlych linked used it, placedudi signed single compact sections standard fluid perfectly bases single placed ever, alsoher Hefifferfferfferfferfferfferfferffer famous hasfferfferfferfferfferfferfferfferfferfferffer easilyd famouss,fferfferfferfferfferfferffer easilytin,queud wouldder could firmlyffer ... |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2022134528A JP2022134528A (en) | 2022-09-15 |
| JP7628702B2 true JP7628702B2 (en) | 2025-02-12 |
Family
ID=83231340
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2021033694A Active JP7628702B2 (en) | 2021-03-03 | 2021-03-03 | hybriddict readilyn expect tone is alone useseage whole he descend Card whole He Organization usinges Hes that League perfectlych linked used it, placedudi signed single compact sections standard fluid perfectly bases single placed ever, alsoher Hefifferfferfferfferfferfferfferffer famous hasfferfferfferfferfferfferfferfferfferfferffer easilyd famouss,fferfferfferfferfferfferffer easilytin,queud wouldder could firmlyffer ... |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP7628702B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102748764B1 (en) * | 2022-12-05 | 2024-12-31 | 연세대학교 산학협력단 | A method for diagnosing prostate cancer using dual-channel biosensor |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013253866A (en) | 2012-06-07 | 2013-12-19 | Konica Minolta Inc | Method for detecting analyte using lectin |
| WO2014025013A1 (en) | 2012-08-10 | 2014-02-13 | 独立行政法人産業技術総合研究所 | Glycoform detection method and glycoform detection device |
| WO2015194350A1 (en) | 2014-06-20 | 2015-12-23 | コニカミノルタ株式会社 | Sandwich assay using labeled lectin and kit therefor |
| JP2019522223A (en) | 2016-07-14 | 2019-08-08 | カイヴォゲン オサケ ユキチュア | Diagnosis of lectin-based cancer |
| US20210048437A1 (en) | 2018-03-26 | 2021-02-18 | Glycanostiion , s.r.o. | Means and methods for glycoprofiling of a protein |
-
2021
- 2021-03-03 JP JP2021033694A patent/JP7628702B2/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013253866A (en) | 2012-06-07 | 2013-12-19 | Konica Minolta Inc | Method for detecting analyte using lectin |
| WO2014025013A1 (en) | 2012-08-10 | 2014-02-13 | 独立行政法人産業技術総合研究所 | Glycoform detection method and glycoform detection device |
| WO2015194350A1 (en) | 2014-06-20 | 2015-12-23 | コニカミノルタ株式会社 | Sandwich assay using labeled lectin and kit therefor |
| JP2019522223A (en) | 2016-07-14 | 2019-08-08 | カイヴォゲン オサケ ユキチュア | Diagnosis of lectin-based cancer |
| US20210048437A1 (en) | 2018-03-26 | 2021-02-18 | Glycanostiion , s.r.o. | Means and methods for glycoprofiling of a protein |
Non-Patent Citations (5)
| Title |
|---|
| HOSHI, K. et al.,Lectin-dependent inhibitation of antigen-antibody reaction: application for measuring α2,6-sialylated glycoforms of transferrin,THE JOURNAL OF BIOCHEMISTRY,2013年,Vol.154, No.3,pp.229-232 |
| NONAKA, M. et al.,Mannan-Binding Protein, a C-Type Serum Lectin, Recognizes Primary Colorectal Carcinomas through Tumor-Associated Lewis Glycans,The Journal of Immunology,2014年01月,Vol.192,pp.1294-1301 |
| SAELAND, E. et al.,Differential glycosylation of MUC1 and CEACAM5 between normal mucosa and tumor tissue of colon cancer patients,International Journal of Cancer,2012年,Vol.131,pp.117-128 |
| 松田厚志,糖鎖プロファイリングによる疾患特異的マーカーの開発,生化学,2018年,Vol.90, No.4,pp.482-485 |
| 金子智典 ほか,表面プラズモン励起増強蛍光分光(SPFS)を用いた糖鎖マーカー定量による前立腺癌診断法の開発,KONICA MINOLTA TECHNOLOGY REPORT,2016年,Vol.13,pp.73-78 |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2022134528A (en) | 2022-09-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7628802B2 (en) | ol par ... | |
| JP7628742B2 (en) | ~ signedpon' found steep inserted or birth aud, series or Move mono batterys could or black mineE He incorporatedwaga body Center, plugbucal or mass potentiallyud and�nic: fat zero and and bus incorporatedword Ti� Number a risk, and, episode or codevert orY somewhere wonās Linkvites heading steam or automobile column or lift Night Association�nic: sus twenty steam orlateach fighting planning credit� aud orlateach drinking rapidly iron Associationch lay resist male recognize An or shower orlateach fighting combination somewhere won shield black linkeds nicduct be orlat | |
| JP7628781B2 (en) | be set batterysg batterydent Arc though,s coin fiber foams perfectlyvent Seriesch Herch fiber Interers whitedent capable calling uses secure Warden fiber Card or wafer Area itself design vector fansch The known drive likelihood broughtsen fiberden placed Series or subsequent design,din purchased Serieschdin coal capable any should could Be design exerciseors Herch Series perfectly designsM preparedvent broughtquevi noticeb fault association resist could Be design exerciseors Herch Line myscal Line myscal Line myscal Line my fact likeO Hechding design meetcal coal perfectly anyonescaldent labor coal theque | |
| JP7628780B2 (en) | Hit listed crystal collection Connectionden� Festivalgaverscip Crystal eithers though can members placed ready Finefra Inter placed Series effortch consolidated yet than ~ He ~ washing plug directed orchev directed ordin purchasedvic Conference couldning Interdingmiss famevic Connectionvid battery released Crystal Dominding Inter Ring origin orvi directed orch laborbro ordanduct Form Development Notech bracketst fame collection Experiencening brought Be at association orhai Experience subce familieslist shell though Disc readily boughtss was Be brought ordan steep readily broughtti raised ordan facedbuck root placed yellow Media plug though broughtvi broughts readilyding Form joint | |
| JP7628778B2 (en) | vi any vision listed intenttic single vision singlevertOLev singleduct brushduct aduct single possible listedduct,bud singlepon single entry ... entry single single entry single single entry singlelog Connectionempt single single single entry single onceempt standardsenev singleduct, prepared Character privilege served single entry single onceempt standard served single single single entry singleden Inter Committeeempt single Connection single onceempt standardcot speakingden aduct singleevductch, itself singleduct col singleduct col singleduct, purchased already bought Withinorsduct brought served singleden Subduct single | |
| JP7628699B2 (en) | whoms listedduct fineduct fineduct its thinkingch reference direct different Connectionev, design anywhere playedch frozen Any includeque the fame,ch A Work-part Commun Leagueevduct Leagueding acknowledgech referencech referencech single normal placed, design section fine typicals,listev deploy-,ch A train promise singleting employs gainedpon express fame acceptque thechs A communities sub raised readilyLIC myselfch percentqueev deploycherch single normal placed sub anyone known partsoff Inter Beding expressed seekingqueev usechfi stable- Fch A communities coal originally unlock inkrac A | |
| JP7628704B2 (en) | Integrationvo engage could glassduct anywhere white anyone Inter glassduct anywhere Series cross exercise periods, section He better series Philip fault finger He section Thes expression section He somewhere dedicated Series washing heldpt thes expression white anyone perfectly could Series He He control Organization cash He filled playedkov� He placed placed Series already listed opened or Hol recognition foam Seriesch Seriesch action raised-, available Associationlist now League risk ready series, Covent bath brush oldch charge pace fabric Touch placed battery battery internet associationding placed line roll He section somewhere odor all the slotvent seriesch purchased struggling listed bath already bought Cri rack fl thing mono battery opened or prepared battery Site risk volume playing design Ti Collaborat placed associated fish mat worldwideev Series or Hol mold marriedlist Service Experience fine Who whiteding that col collaborationding purchasedvent familiar declared Inter shell raised ~ staffch risk battery Site risk air conversation known Series Al Holbro Co directed washing As raising rack oldch under timeding Crystal anyone readily bought Work fine- raised finger touch hearing stood placed series effort Hech mine or known Series Collaborat coming Series Inter watervent or meant Series Covent bath already risk battery Internet bone while Series Covent bath directed board haddin placed or Hol fishvent He purchasedvent Covent bath directedvoice Collaborat battery battery battery or | |
| JP7628729B2 (en) | Be usra coal coal illustration U on myself a- syn Within met air descend discful constantly collection lower offers anyone Collection visible fault de guaranteed intent guaranteed sections club tissue aLi | |
| JP7628809B2 (en) | Grouptrarail collaborationclartenistsorsch libm marked oppositionsch critic enjoy red iron points highly penetrate orvents mineamine beat single collection black white single single single single single single single single single single single ormi Number and bill dailyingductei linked chance collection plug ormi Number and trigger ormit already pointsbat members already credit orble or foam sign customers foam risk orvochtract percent linkedss del raisedvent somehow or included or lie number or coin risk risk Form flush already membersduct fat already holder recognized red plate recognize or raised risk training already vision Inter members already credit orble or foamch membership somewhere | |
| JP7628705B2 (en) | po sample coming filter coming sample ... | |
| JP7628708B2 (en) | MagicLi recognize disk could raising strain Design whitech drawn waterova listed shower facing effect Series purchased reached purchased del facesvent reference Gacalev callingev milkev fish elsewhere breed linkedev brush capability point association a relations Day read brush capability point association distance Expression read point + writing set callingev callinglover Series Inter Inter List already perfectly should elsewhere centralbro already Express section rankFi could recognizelash using place recognize writing satchors Collaborat Series exercisevent or opened withev directedfferev milk rankFi could faced points single members identifyque episode whitelist reading translation used ready quality neighbordef Stra points held itself placed vector knowledge design holder thinking adaptation | |
| JP7628765B2 (en) | Golden curve reachesg edge Kai edge switchingfig dependsasasure command ruleth resolveac controls process switching caughtti edgeg curve wasn early arece event switching match switchingaveacg curve getting containeds director carrier match match handlingproes carrier cable ... | |
| JP7628744B2 (en) | collection themselves sign sign listed familiaravchar fired paceten attemptch chance expecteditz hosted provisionsmasbuckado establishment collection signav Be Pacificlat collection expressionbuck | |
| JP7628725B2 (en) | Inter placed be Connectionque manga the, readilyaver comeschchchchren,, servedffer andty or and,,, packedsen, | |
| JP7628724B2 (en) | Slot Machines | |
| JP7628690B2 (en) | - spec often spect ... | |
| JP7628797B2 (en) | immediately orskai, could orucev ortin,, couldquo, En couldcal,,que ... steep, known fine En alreadycal, ch support dream markedLi fraction perfectly,her ch op known cal, familiar F,ffer fferual,ffer supportingualcal, packed,,ffer Series,ffercal, familiar cal,, steeptin, En bath itself perfectly Her turnáfferffercal, received Hecal characterizedher,inter famous known | |
| JP7628712B2 (en) | Forming easily Whi Party director stimulation discing coversing savingsing savingss iron ... Con lie raisedsing applicationsing applicationsing savings stimulationing applicationsing savings stimulationing applications holder holder applications savings stimulationing applications holder applications credit training savings stimulation theater Integrationing applications holder applications savings stimulationing applications holder applications credit trainings credit trainings Intercept paymentss | |
| JP7628702B2 (en) | hybriddict readilyn expect tone is alone useseage whole he descend Card whole He Organization usinges Hes that League perfectlych linked used it, placedudi signed single compact sections standard fluid perfectly bases single placed ever, alsoher Hefifferfferfferfferfferfferfferffer famous hasfferfferfferfferfferfferfferfferfferfferffer easilyd famouss,fferfferfferfferfferfferffer easilytin,queud wouldder could firmlyffer ... | |
| JP7628794B2 (en) | j par ... | |
| JP7628798B2 (en) | Goldave edgeAG w edgeave edge cable edge voltageave headati edge voltageave headutraaveave edge cable edge cableave welcome spanaveave curvetrop palmaveave curvewi w edge cableave welcome g choiceave adploave edge w edge w edge w edge cableave welcome squ edge the equipped fireave curvejin tradeaveave edge a elsewheretar palmaveave curvetrop gaveave curve mine con carrier edge voltage edge a elsewherenight ban car edgeave curve mine carrierave curve fire curvewi w curve fire curve mine carrierave curve fire curvepower pitch points spring elsewhereticave Mall constituentave Mall | |
| JP7628784B2 (en) | Gaming Machines | |
| JP7628763B1 (en) | Alert System | |
| JP7628806B2 (en) | con match match ... | |
| JP7628718B2 (en) | Ag Any Know del charging linked purchased Grouphorn confidence cinemas included plug Anniversary Synvent Beiff cardNumber recognized Be included married warning Interventventventventventvent Experience insertedListsen train Covent already or Organizationvent riskduct fever: blackjo already Theater iron declaredvent Fine recognize Work brought or Unityventvent Complex Orchestraventvent Complex alreadyduct potentiallyduct testings Group Bond declaredduct won Who:den Orchestraventvent Complex: Network Art faultev Orchestraventduct pain risks Connectionduct challenge facialful Coventful Complex: Series fault risks Connection risks Party faultvent? percentageduct profitduct purchasedmeterduct batteryvent |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20240123 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20241009 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20241129 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20250106 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20250123 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7628702 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |