JPH0122905B2 - - Google Patents
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- Publication number
- JPH0122905B2 JPH0122905B2 JP56015013A JP1501381A JPH0122905B2 JP H0122905 B2 JPH0122905 B2 JP H0122905B2 JP 56015013 A JP56015013 A JP 56015013A JP 1501381 A JP1501381 A JP 1501381A JP H0122905 B2 JPH0122905 B2 JP H0122905B2
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- Prior art keywords
- antigen
- antibody
- enzyme
- immunoglobulin
- column
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/537—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody
- G01N33/538—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody by sorbent column, particles or resin strip, i.e. sorbent materials
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- Engineering & Computer Science (AREA)
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- Biomedical Technology (AREA)
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- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】
本発明は抗原を速かにかつ精度良く測定する方
法に関するものである。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for rapidly and accurately measuring antigens.
更に詳しくは抗原を定量するに際して、定量す
べき抗原と酵素標識抗原とを該抗原に対する抗体
イムノグロブリンに対して競争的あるいは拮抗的
に反応させ、得られた反応液を、第二抗体不溶化
担体に接触せしめ、反応によつて生成した酵素標
識抗原と抗体イムノグロブリンの結合物を第二抗
体不溶化担体に吸着せしめ、第二抗体不溶化担体
に吸着されずに液相に残存する遊離の酵素標識抗
原を定量することによつて、抗原を速かにかつ精
度良く測定することを基本的技術思想とする抗原
の定量法に関するものである。 More specifically, when quantifying an antigen, the antigen to be quantified and the enzyme-labeled antigen are reacted competitively or competitively with the antibody immunoglobulin against the antigen, and the resulting reaction solution is transferred to a second antibody-insolubilized carrier. The conjugate of the enzyme-labeled antigen and antibody immunoglobulin produced by the reaction is adsorbed onto the second antibody-insolubilized carrier, and the free enzyme-labeled antigen remaining in the liquid phase without being adsorbed to the second antibody-insolubilized carrier is removed. This invention relates to a method for quantifying antigens whose basic technical idea is to rapidly and accurately measure antigens through quantitative determination.
本発明において定量される抗原としては種々の
生体成分あるいは薬剤があるが、特に臨床的には
生体体液中に含まれる種々のホルモン類、蛋白質
が重要である。具体的にはペプチドホルモン類、
ステロイドホルモン類、サイロキシン（T4）、ト
リヨードチロニン（T3）等の甲状線ホルモンあ
るいはイムノグロブリン、α−フエトプロテイン
（AFP）、癌胎児性抗原（CEA）等がある。 Antigens to be quantified in the present invention include various biological components or drugs, and in particular clinically important are various hormones and proteins contained in biological body fluids. Specifically, peptide hormones,
Examples include steroid hormones, thyroid hormones such as thyroxine (T 4 ) and triiodothyronine (T 3 ), immunoglobulins, α-fetoprotein (AFP), and carcinoembryonic antigen (CEA).
一般に免疫測定法は測定感度あるいは特異性に
おいて優れており生体体液中の微量物質の定量に
広く用いられる方法であつて、この方法には標識
物質の種類によりラジオイムノアツセイ（RIA）、
蛍光免疫測定法（FIA）、酵素免疫測定法（EIA）
等に分類される。 In general, immunoassays are excellent in measurement sensitivity and specificity, and are widely used for quantifying trace substances in biological body fluids.This method includes radioimmunoassay (RIA),
Fluorescence immunoassay (FIA), enzyme immunoassay (EIA)
etc.
このうち、ラジオイムノアツセイは自動測定機
器が開発され広く普及しているが、特殊な施設と
技術者あるいは管理者を必要とし、また標識物質
として放射性同位元素を用いるため測定廃棄物の
処理に厳重な注意を要する等の問題がある。 Among these, radioimmunoassay has developed automatic measuring equipment and is widely used, but it requires special facilities and technicians or managers, and because it uses radioactive isotopes as labeling substances, it is difficult to dispose of measurement waste. There are problems that require strict attention.
一方、酵素免疫測定法においては、これらの問
題点は解決されるが、いわゆる固相法においては
不溶性担体上に結合した酵素活性を測定するた
め、固相（不溶性担体）の洗浄等煩雑な操作を必
要とし、自動測定系への導入も難しく、また生体
体液成分による干渉を受けて測定精度が低いとい
う欠点がある。 On the other hand, in enzyme immunoassay, these problems are solved, but in the so-called solid-phase method, the enzyme activity bound on an insoluble carrier is measured, so complicated operations such as washing the solid phase (insoluble carrier) are required. It is difficult to introduce into an automatic measurement system, and the measurement accuracy is low due to interference from biological fluid components.
本発明者等は、以上の点を解決すべく鋭意検討
した結果、第二抗体不溶化担体のカラムを用いて
遊離の酵素標識抗原を定量することにより速かに
かつ精度良く抗原を測定でき、しかも自動測定系
への導入が極めて容易であるところの抗原の定量
法を確立したのである。 As a result of intensive studies to solve the above points, the present inventors have found that by quantifying free enzyme-labeled antigen using a column of a second antibody-insolubilized carrier, it is possible to quickly and accurately measure the antigen. They established a method for quantifying antigens that is extremely easy to introduce into an automatic measurement system.
即ち、本発明は、定量すべき抗原と酵素標識抗
原を該抗原に対する抗体イムノグロブリンに競争
的に反応させ、得られた反応液を第二抗体不溶化
担体のカラムに流し、反応によつて生成した酵素
標識抗原と抗体イムノグロブリンの結合物を該反
応液より除いた後、液相（カラムからの流下液）
の残存する遊離の酵素標識抗原量を、酵素活性を
測定することによつて求め、その測定値より抗原
を定量する方法であり、しかも上記工程における
抗イムノグロブリン抗体と不溶性担体との結合
は、Ｓ−Ｓ結合を介して行い、使用後、該不溶性
担体を還元剤にて洗浄し、再び抗イムノグロブリ
ン抗体をＳ−Ｓ結合を介して結合せしめ、くり返
し測定に使用するものであり、且つ測定系には、
ゼラチンと塩類とを共存させて、生体体液中に含
まれる干渉物質の影響を抑制しつつ定量する方法
である。 That is, in the present invention, an antigen to be quantified and an enzyme-labeled antigen are competitively reacted with an antibody immunoglobulin against the antigen, and the resulting reaction solution is flowed through a column of a second antibody-insolubilized carrier, and the antibody produced by the reaction is After removing the conjugate of enzyme-labeled antigen and antibody immunoglobulin from the reaction solution, the liquid phase (flowing liquid from the column)
The remaining free enzyme-labeled antigen amount is determined by measuring the enzyme activity, and the antigen is quantified from the measured value, and the binding of the anti-immunoglobulin antibody and the insoluble carrier in the above step is After use, the insoluble carrier is washed with a reducing agent, and the anti-immunoglobulin antibody is bound again via the S-S bond to be used for repeated measurements. In the system,
This is a method in which gelatin and salts coexist to quantify the amount while suppressing the influence of interfering substances contained in biological body fluids.
本発明に使用される標識用酵素としてはβ−ガ
ラクトシダーゼ、アルカリホスフアターゼ等EIA
において通常用いられる酵素であればいずれでも
よい。 Labeling enzymes used in the present invention include β-galactosidase, alkaline phosphatase, etc.
Any enzyme commonly used can be used.
酵素標識抗原の作製に際してはグルタルアルデ
ヒド、カルボジイミド、Ｎ，Ｎ−ｏ−フエニレン
ジマレイミド、ｍ−マレイミドベンゾイル−Ｎ−
ハイドロキシサクシンイミドエステル（ｍ−
Maleimidobenzoyl−Ｎ−Hydroxysuccinimide
Ester）等の二官能性試薬が用いられる。 When preparing an enzyme-labeled antigen, glutaraldehyde, carbodiimide, N,N-o-phenylene dimaleimide, m-maleimidobenzoyl-N-
Hydroxysuccinimide ester (m-
Maleimidobenzoyl-N-Hydroxysuccinimide
Bifunctional reagents such as Ester) are used.
不溶性担体としてはアガロース、デキストラ
ン、セルロースなどの多糖類、ポリエチレン等の
合成樹脂あるいはガラス、ポリアクリルアミド等
が用いられ、形態としてはビーズ状、繊維状であ
ることが好ましい。 As the insoluble carrier, polysaccharides such as agarose, dextran, and cellulose, synthetic resins such as polyethylene, glass, polyacrylamide, etc. are used, and the form is preferably beads or fibers.
抗イムノグロブリン抗体と不溶性担体との結合
は、Ｓ−Ｓ結合を利用して行い、例えば抗イムノ
グロブリン抗体の分子に存在するＳ−Ｓ結合を還
元し、あるいはＳ−アセチルメルカプト無水コハ
ク酸等の試薬を用いてSH基を導入し、これとSH
基を有する不溶性担体とを接触せしめてＳ−Ｓ結
合によつて不溶化せしめる。この場合、測定に使
用した不溶性担体をジチオスレイトールの如き還
元剤で洗浄し、不溶性担体上のＳ−Ｓ結合を切断
してSH基とし、再びSH基を付与せしめた抗イム
ノグロブリンを結合させ、くり返し測定に使用す
る。 The binding between the anti-immunoglobulin antibody and the insoluble carrier is carried out using an S-S bond, for example, by reducing the S-S bond present in the anti-immunoglobulin antibody molecule, or by using S-acetylmercaptosuccinic anhydride or the like. A reagent is used to introduce an SH group, and this and SH
It is brought into contact with an insoluble carrier having a group and insolubilized by S--S bonds. In this case, the insoluble carrier used in the measurement is washed with a reducing agent such as dithiothreitol, the S-S bond on the insoluble carrier is cleaved to form an SH group, and the anti-immunoglobulin to which the SH group has been added is bound again. , used for repeated measurements.
ここで使用される抗イムノグロブリン抗体は、
イムノグロブリンそのものでもよいが、ペプシ
ン、パパインなどのプロテアーゼで処理して抗原
結合部位のみを分離したもの（即ちFab′部分、
Ｆ（ab′）2部分など）であつてもよい。 The anti-immunoglobulin antibody used here is
Immunoglobulin itself may be used, but it may be treated with proteases such as pepsin or papain to isolate only the antigen-binding site (i.e., Fab' portion,
F(ab′) 2 parts, etc.).
上のような試薬を用いることにより抗原の定量
を行なうことができるが、生体体液中の微量成分
を測定するような場合には、生体体液成分による
干渉作用により測定精度が著しく悪くなることが
あるので、測定系にゼラチンと塩類を共存させる
ことにより、干渉物質による影響を抑制あるいは
除去する。塩類としては食塩等が用いられる。 Antigens can be quantified by using the above reagents, but when measuring trace amounts of components in biological fluids, measurement accuracy may deteriorate significantly due to interference from biological fluid components. Therefore, by coexisting gelatin and salts in the measurement system, the influence of interfering substances can be suppressed or eliminated. Common salt or the like is used as the salt.
本発明によればこのように、簡単な操作で精度
良く抗原を測定することができ、また酵素活性を
測定する試料が溶液であるため自動測定系への応
用が可能となるのである。 According to the present invention, antigens can be measured with high precision using simple operations, and since the sample for measuring enzyme activity is a solution, it is possible to apply the method to an automatic measurement system.
次に本発明の実施例を示す。 Next, examples of the present invention will be shown.
実施例 １
サイロキシン（T4）の定量
(1) ヤギ（抗ウサギIgG）IgGの調製（IgG＝イ
ムノグロブリンＧ）
加藤らの方法（Journal of Biochemistry
81巻、1557ページ、1977年）に従い、ヤギの抗
ウサギIgG血清より硫安分画、DEAE−セルロ
ースクロマトグラフイによつて調製した。Example 1 Quantification of thyroxine (T 4 ) (1) Preparation of goat (anti-rabbit IgG) IgG (IgG = immunoglobulin G) Method of Kato et al. (Journal of Biochemistry
81, p. 1557, 1977), it was prepared from goat anti-rabbit IgG serum by ammonium sulfate fractionation and DEAE-cellulose chromatography.
(2) 再生可能な（抗ウサギIgG）IgG不溶化セフ
アロースの調製
(1)で調製した（抗ウサギIgG）IgG溶液を
0.1M酢酸緩衝液（PH５）に対して透析し、つ
いで75ｍＭの２−メルカプトエチルアミンにて
37℃で90分間還元した。得られた還元IgGは１
分子当り６〜７個のSH基を持つていた。次に
0.1M酢酸緩衝液（PH５）で平衡化したセフア
デツクスＧ−25のカラムで２−メルカプトエチ
ルアミンを除き、同じ緩衝液で平衡化したセフ
アロース−（グルタチオン−２−ピリジルジス
ルフイド）のカラムにこの還元IgGを流し、
IgGをＳ−Ｓ結合を介してセフアロースに不溶
化した。（抗ウサギIgG）IgG不溶化セフアロー
スは0.3M KClと１ｍＭエチレンジアミン４酢
酸を含む0.1Mトリス塩酸緩衝液（PH7.5）で洗
浄後４℃で保存した。(2) Preparation of reproducible (anti-rabbit IgG) IgG insolubilized sepharose The (anti-rabbit IgG) IgG solution prepared in (1)
Dialyzed against 0.1M acetate buffer (PH5), then 75mM 2-mercaptoethylamine.
Reduction was performed at 37°C for 90 minutes. The obtained reduced IgG was 1
It had 6-7 SH groups per molecule. next
The 2-mercaptoethylamine was removed on a column of Sephadex G-25 equilibrated with 0.1 M acetate buffer (PH5), and this reduction was transferred to a column of Sepharose (glutathione-2-pyridyl disulfide) equilibrated with the same buffer. flowing IgG,
IgG was insolubilized in Sepharose via SS bonds. (Anti-rabbit IgG) IgG-insolubilized Sepharose was washed with 0.1M Tris-HCl buffer (PH7.5) containing 0.3M KCl and 1mM ethylenediaminetetraacetic acid, and then stored at 4°C.
(3) β−ガラクトシーゼ標識サイロキシンの調製
サイロキシンと４−（マレイミドメチル）シ
クロヘキサン−１−カルボキシリツクアシツド
サクシンイミドエステル（４−
（Maleimidomethyl）cyclohexane−１−
carboxylic acid，succinimide ester、
MCAE、ジーベンケミカル社、東京）を各々
別にジメチルホルムアミドに溶解し、0.1Mリ
ン酸緩衝液（PH７）に加えて混合し、30℃で90
分反応させた。反応は1Mグリシン溶液を加え
ることによつて停止した。次にこの反応液に
0.1Mリン酸緩衝液（PH７）に溶解したβ−ガ
ラクトシダーゼを加え、30℃で30分間反応させ
た後、0.1Mメルカプトエチルアミンを加えて
反応を停止し、この反応液を0.1Mリン酸緩衝
液（PH７）で平衡化したセフアクリルＳ−300
（フアルマシア社）のカラムに流し、活性ピー
クの部分を集めた。得られたβ−ガラクトシダ
ーゼ標識サイロキシンは、塩化マグネシウムと
牛血清アルブミンを加えて４℃で保存した。(3) Preparation of β-galactosase-labeled thyroxine Thyroxine and 4-(maleimidomethyl)cyclohexane-1-carboxylic acid succinimide ester (4-
(Maleimidomethyl)cyclohexane-1-
carboxylic acid, succinimide ester,
MCAE, Sieben Chemical Co., Tokyo) were each dissolved in dimethylformamide separately, added to 0.1M phosphate buffer (PH7), mixed, and incubated at 30℃ for 90 minutes.
It was allowed to react for a minute. The reaction was stopped by adding 1M glycine solution. Next, add this reaction solution to
β-galactosidase dissolved in 0.1M phosphate buffer (PH7) was added and reacted at 30°C for 30 minutes, then 0.1M mercaptoethylamine was added to stop the reaction, and the reaction solution was added to 0.1M phosphate buffer. Cephacryl S-300 equilibrated with (PH7)
(Pharmacia) column, and the active peak portion was collected. The obtained β-galactosidase-labeled thyroxine was stored at 4°C with the addition of magnesium chloride and bovine serum albumin.
(4) β−ガラクトシターゼ活性の測定
酵素液（PH７）１mlにｏ−ニトロフエニル−
β−Ｄ−ガラクトピラノシド溶液（4.5mg／ml）
0.25mlを加え、37℃で反応させた液、1M炭酸
ナトリウム溶液0.25mlを加えて反応を停止し、
420nｍの吸光度を測定した。(4) Measurement of β-galactosidase activity Add 1 ml of enzyme solution (PH7) to o-nitrophenyl-
β-D-galactopyranoside solution (4.5mg/ml)
Add 0.25ml of the solution and react at 37℃, add 0.25ml of 1M sodium carbonate solution to stop the reaction,
Absorbance at 420 nm was measured.
(5) 測定
標準サイロキシン溶液を緩衝液Ｇ（0.3M
NaCl、１ｍＭ MgCl2、0.5％ゼラチン、0.1％
牛血清アルブミン、0.1％NaN3 ２ｍＭ Ｎ−
エチルマレイミドを含むPH７の10ｍＭリン酸緩
衝液）にて希釈した各検体0.1ml（サイロキシ
ン変量０、0.1、0.3、1.0、3.0、10、30nｇ／0.1
ml）、緩衝液Ｇで100倍に希釈した抗サイロキシ
ン血清（ウサギ）0.1mlを混合し、更にβ−ガ
ラクトシダーゼ標識サイロキシンを含む緩衝液
G0.3mlを加え、30℃で１時間反応させた。こ
の反応液を緩衝液Ｇで平衡化した（抗ウサギ
IgG）IgG不溶化セフアロースのカラム（0.1
ml）に流した後、カラムを２mlの緩衝液Ｇで洗
浄した。カラムより流下した液を集め、その酵
素活性を測定してサイロキシン検量線を得た。
なお、免疫反応において標準サイロキシンと抗
サイロキシン血清を１時間反応させた後、β−
ガラクトシターゼ標準サイロキシンを加え更に
１時間反応させた場合も同じ検量線が得られ
た。また１時間の反応は少なくとも15分まで短
縮しても測定に支障はなかつた。(5) Measurement Standard thyroxine solution was added to buffer G (0.3M
NaCl, 1mM MgCl2 , 0.5% gelatin, 0.1%
Bovine serum albumin, 0.1% NaN 3 2mM N-
0.1ml of each sample diluted with 10mM phosphate buffer (PH7 containing ethylmaleimide) (thyroxine variation 0, 0.1, 0.3, 1.0, 3.0, 10, 30ng/0.1
ml), mix 0.1 ml of anti-thyroxine serum (rabbit) diluted 100 times with buffer G, and add buffer containing β-galactosidase-labeled thyroxine.
0.3 ml of G was added and reacted at 30°C for 1 hour. This reaction solution was equilibrated with buffer G (anti-rabbit
IgG) IgG insolubilized sepharose column (0.1
ml), the column was washed with 2 ml of buffer G. The liquid flowing down from the column was collected and its enzyme activity was measured to obtain a thyroxine calibration curve.
In addition, in the immune reaction, after reacting standard thyroxine and anti-thyroxine serum for 1 hour, β-
The same calibration curve was obtained when the galactosidase standard thyroxine was added and reacted for an additional hour. Furthermore, the 1-hour reaction time could be shortened to at least 15 minutes without any problem in measurement.
実施例 ２
不溶性担体の再生
実施例１において使用した（抗ウサギIgG）
IgG不溶化セフアロースを再生した。Example 2 Regeneration of insoluble carrier Used in Example 1 (anti-rabbit IgG)
IgG insolubilized sepharose was regenerated.
測定に使用したカラム（0.1ml）を0.3M KClと
１ｍＭエチレンジアミン４酢酸を含むPH7.5の
0.1Mトリス塩酸緩衝液で洗浄し、次に30ｍＭの
ジチオスレイトールを含む同じ緩衝液を流し、更
にカラムを洗浄後、２−2′−ジチオジピリジンを
含む緩衝液を流してＳ−Ｓ結合を形成させた。こ
のカラムに実施例１と同様の方法で（抗ウサギ
IgG）IgGを結合させた。 The column (0.1ml) used for measurement was heated to pH 7.5 containing 0.3M KCl and 1mM ethylenediaminetetraacetic acid.
Wash the column with 0.1M Tris-HCl buffer, then run the same buffer containing 30mM dithiothreitol, wash the column again, and run a buffer containing 2-2'-dithiodipyridine to complete the S-S bond. formed. This column was coated in the same manner as in Example 1 (anti-rabbit
IgG) IgG was bound.
再生したカラムを用いてサイロキシンを定量し
たところ実施例１の場合と同じ検量線が得られ
た。 When thyroxine was quantified using the regenerated column, the same calibration curve as in Example 1 was obtained.
Claims (1)
素標識抗原）とを該抗原に対する抗体イムノグロ
ブリンに競争的に反応させ； 得られた反応液を、抗イムノグロブリン抗体を
結合せしめた不溶性担体（第二抗体不溶化担体）
のカラムに流し、該反応液中の酵素標識抗原と抗
体イムノグロブリンの結合物を該カラムに吸着せ
しめ； しかも、上記工程における抗イムノグロブリン
抗体と不溶性担体との結合はＳ−Ｓ結合を介して
行い、使用後、該不溶性担体を還元剤にて洗浄
し、再び抗イムノグロブリン抗体をＳ−Ｓ結合を
介して結合せしめ、くり返し測定に使用するもの
であり； 且つ測定系には、ゼラチンと塩類とを共存させ
て、生体体液中に含まれる干渉物質の影響を抑制
せしめ； 上記吸着工程後、該カラムに吸着されずに流下
する遊離の酵素標識抗原を定量すること； を特徴とする酵素標識抗原と第二抗体不溶化担体
を用いる抗原の定量法。[Claims] 1. The antigen to be quantified and the antigen labeled with an enzyme (enzyme-labeled antigen) are competitively reacted with an antibody immunoglobulin against the antigen; the resulting reaction solution is treated with an anti-immunoglobulin antibody. Bound insoluble carrier (second antibody insolubilized carrier)
The conjugate of the enzyme-labeled antigen and antibody immunoglobulin in the reaction solution is adsorbed onto the column; Moreover, the binding between the anti-immunoglobulin antibody and the insoluble carrier in the above step is via an S-S bond. After use, the insoluble carrier is washed with a reducing agent, and the anti-immunoglobulin antibody is bound again via an S-S bond, and used for repeated measurements; and the measurement system includes gelatin and salts. suppressing the influence of interfering substances contained in biological body fluids; quantifying the free enzyme-labeled antigen flowing down without being adsorbed to the column after the adsorption step; Antigen quantification method using antigen and second antibody insolubilized carrier.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1501381A JPS57131062A (en) | 1981-02-05 | 1981-02-05 | Quantitative determination of antigen using enzyme- labelled antigen and second antibody insolubilizing carrier |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1501381A JPS57131062A (en) | 1981-02-05 | 1981-02-05 | Quantitative determination of antigen using enzyme- labelled antigen and second antibody insolubilizing carrier |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS57131062A JPS57131062A (en) | 1982-08-13 |
| JPH0122905B2 true JPH0122905B2 (en) | 1989-04-28 |
Family
ID=11876988
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1501381A Granted JPS57131062A (en) | 1981-02-05 | 1981-02-05 | Quantitative determination of antigen using enzyme- labelled antigen and second antibody insolubilizing carrier |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS57131062A (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3617763A1 (en) * | 1985-05-28 | 1986-12-04 | Olympus Optical Co., Ltd., Tokio/Tokyo | METHOD FOR CARRYING OUT IMMUNOLOGICAL PROVISIONS AND APPARATUS APPARATUS FOR THIS |
| US4810638A (en) * | 1986-07-24 | 1989-03-07 | Miles Inc. | Enzyme-labeled antibody reagent with polyalkyleneglycol linking group |
| JP2638521B2 (en) * | 1994-11-24 | 1997-08-06 | 有限会社ビー・エス・アール | Determination of liver disease by measurement of serum bile acids by ELISA |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1036935A (en) * | 1973-03-19 | 1978-08-22 | Lavell R. Johnson | Method and apparatus for radioimmunoassay with regeneration of immunadsorbent |
| ZA761751B (en) * | 1975-04-07 | 1977-03-30 | Summa Corp | Immobilized immunoadsorbent |
| US4128628A (en) * | 1976-03-12 | 1978-12-05 | University Of Virginia Alumni Patents Foundation | Automated immunoassay |
| US4104026A (en) * | 1976-03-12 | 1978-08-01 | University Of Virginia | Immunoassay separation technique |
| JPS5925184B2 (en) * | 1978-12-22 | 1984-06-15 | 天野製薬株式会社 | Method for eliminating nonspecific inhibitory effects in immunoassays |
| JPS55136261A (en) * | 1979-04-12 | 1980-10-23 | Toyo Jozo Co Ltd | Novel compound and kit comprising it |
-
1981
- 1981-02-05 JP JP1501381A patent/JPS57131062A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS57131062A (en) | 1982-08-13 |
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