JP4337643B2 - Biochemical instruments - Google Patents

Description

The present invention relates to a biochemical instrument.

  Conventionally, glass products have been used as test instruments in the life science field that makes full use of culture techniques such as plant tissue culture and animal cell culture and immunobiochemical techniques. However, resin products have come to be used due to the recent increase in hygiene awareness of infections, etc., the lowering of price, higher purity, and advances in resin product molding technology associated with mass production of resin products. Yes.

  Such resin products include disposable biochemical instruments (for example, plates used for immunization experiments such as antigen-antibody reaction quantification using the EIA / ELISA method, structure / function analysis of proteins, and clinical tests). Biochemical containers such as dilution tubes and storage tubes). Such a biochemical instrument is required to have a high adsorption ability with protein depending on the purpose. For example, in the case of a plate used for an immunization experiment, an oxygen plasma treatment, an ultraviolet treatment, etc. are used to adjust the hydrophilic-hydrophobic balance by making the surface of the resin product hydrophilic, thereby increasing the adsorption property to proteins ing.

  Further, in tubes such as blood collection containers or test tubes and sample containers used for protein analysis, it is not preferable that biological substances such as proteins are adsorbed on the surfaces thereof. For example, when protein in urine or blood is adsorbed to a container in a clinical test, the test value for the amount of protein is lower than the actual value or not detected at all, and the diagnosis may be wrong. Also, in protein analysis, especially when dealing with a low concentration of a minute amount of protein, if it is adsorbed to a container in a dilution or storage step, the concentration changes, which greatly affects the analysis result.

  As means for solving such a problem, a method of modifying the surface of a polystyrene member by circulation of ozone gas is disclosed (for example, see Patent Document 1). However, it is known that polar groups introduced on the polystyrene surface will sink into the resin layer over time due to contact with the gas phase, and the treatment effect will decrease over time. It was.

JP-A-10-101820

  The objective of this invention is providing the instrument for biochemistry which can reduce the adsorption amount of a biological substance.

（２）前記生化学用器具の表面には、第２の官能基が形成されているものである上記（１）に記載の生化学用器具。
（３）前記生化学用器具に対する前記水溶性樹脂の固定は、主として前記第１の官能基と前記第２の官能基とが共有結合することにより行なわれているものである上記（２）に記載の生化学用器具。
（４）生体由来物質の吸着率が、２０％以下である上記（１）ないし（３）のいずれかに記載の生化学用器具。
（５）アセトニトリルの５０容量％水溶液を１０分間接触させた後の生体由来物質の吸着率が、２５％以下である上記（１）ないし（４）のいずれかに記載の生化学用器具。
（６）前記生体由来物質は、ウシ血清アルブミンまたはウシ血清イムノグロブリンＧである上記（４）または（５）に記載の生化学用器具。
（７）前記生化学用器具は、チューブとして用いられるものである上記（１）ないし（６）のいずれかに記載の生化学用器具。 Such an object is achieved by the present invention described in the following (1) to (7).
(1) A water-soluble resin having a first functional group on a side chain is fixed to a biochemical instrument composed of a resin material,
The water-soluble resin having the first functional group in the side chain is polyvinyl alcohol obtained by saponifying polyvinyl acetate, and the polyvinyl alcohol further includes a structural unit represented by the following formula (Ia ). Biochemical instrument characterized by being a thing.

(2) The biochemical instrument according to (1) above, wherein a second functional group is formed on the surface of the biochemical instrument.
(3) the fixing of the water-soluble resin to the biochemical instrument is above SL is what is done mainly by the first functional group and the second functional group is covalently attached (2) biochemical instrument according to.
(4) The biochemical instrument according to any one of the above (1) to (3), wherein the adsorption rate of the biological substance is 20% or less.
(5) The biochemical instrument according to any one of the above (1) to (4), wherein the adsorption rate of the biological substance after contacting with a 50% by volume aqueous solution of acetonitrile for 10 minutes is 25% or less.
(6) The biochemical instrument according to (4) or (5), wherein the biological substance is bovine serum albumin or bovine serum immunoglobulin G.
(7) The biochemical instrument according to any one of (1) to (6), wherein the biochemical instrument is used as a tube.

ADVANTAGE OF THE INVENTION According to this invention, the biochemical instrument which can reduce the adsorption amount of a biological substance can be provided.
Further, when the water-soluble resin having the first functional group in the side chain is cured and used, a biochemical instrument that is particularly excellent in heat resistance and that has a particularly small amount of adsorption of a biological substance after heat treatment is provided. be able to.
As described above, according to the present invention, since the adsorption rate of the biological substance is low, it is possible to provide a container for analyzing a biological substance having a low concentration.

Hereinafter, the biochemical instrument of the present invention will be described in detail.
The biochemical instrument of the present invention is characterized in that a water-soluble resin having a first functional group in the side chain is fixed to a biochemical instrument composed of a resin material.

First, the water-soluble resin having the first functional group in the side chain will be described. The water-soluble resin having the first functional group in the side chain is fixed to the biochemical instrument (the surface of the biochemical instrument). Thereby, the adsorption amount of a biological substance can be reduced.
The first functional group preferably includes a photosensitive reactive group, a radiation reactive reactive group, and a thermosensitive reactive group. Among these, it is particularly preferable to contain a photosensitive reactive group. Thereby, the water-soluble resin having the first functional group in the side chain can be cross-linked efficiently in a short time with a simple production facility.

Examples of the first functional group include a functional group containing a nitrogen atom, a functional group containing a sulfur atom, a functional group containing a bromine atom, a functional group containing a chlorine atom, or a functional group containing neither of these atoms. Is mentioned. Among these, a functional group containing a nitrogen atom is preferable.
Specific examples include a functional group containing an azide group, a functional group containing a diazo group, and a functional group containing a diazide group. Among these, a functional group containing an azide group is preferable. Thereby, it can be made to react with the wavelength of practical 300-500 nm, and the formability of a film | membrane can be improved with the further outstanding resolution.

As the first functional group, it is preferable to use one specifically represented by the following formula (II). The functional group can be easily imparted to the water-soluble resin, and at the same time has excellent reactivity at the practical wavelength of 300 to 500 nm, so that a good water-soluble resin film can be formed.

  The modification rate (substitution rate) of the first functional group is preferably from 0.1 to 20 mol%, and preferably from 0.2 to 10 mol% of the entire water-soluble resin having the first functional group in the side chain. If the modification rate is less than the lower limit, the photosensitivity may be lowered, and if it exceeds the upper limit, it may be difficult to form a uniform film.

As the water-soluble resin constituting the water-soluble resin having the first functional group in the side chain, for example, saponified polyvinyl acetate, polyvinyl pyrrolidone, polyethylene glycol, polyacrylamide, polymethacrylamide, polyhydroxy Ethyl methacrylate, polypentaerythritol triacrylate, polypentaerythritol tetraacrylate, polydiethylene glycol diacrylate, and copolymers of monomers constituting them, and 2-methacryloyloxyethyl phosphorylcholine and other monomers (for example, butyl methacrylate) And a copolymer thereof. Among these, one or more selected from saponified products of polyvinyl acetate, polyvinyl pyrrolidone, and polyethylene glycol are preferable. Thereby, the reduction effect of the adsorption amount of a biological substance can be improved.
Here, the polyvinyl acetate fight is a polyvinyl alcohol or a copolymer of vinyl alcohol and another compound. Further, vinyl alcohol and modified vinyl acetate saponified products such as hydrophilic group modification, hydrophobic group modification, anion modification, cation modification, amide group modification or reactive group modification such as acetoacetyl group are included.
In addition, water-soluble resin means what can melt | dissolve 1.0g or more with respect to 100g of 25 degreeC water here.

  The average degree of polymerization of the water-soluble resin is not particularly limited, but is preferably 100 to 10,000, and particularly preferably 200 to 5,000. If the average degree of polymerization is less than the lower limit, it may be difficult to uniformly form a film on the surface of the biochemical instrument, and if the upper limit is exceeded, the side chain has the first functional group. In some cases, the viscosity of the water-soluble resin increases and the workability decreases.

  Further, when the saponified product of polyvinyl acetate is used, the saponification degree of the saponified product of polyvinyl acetate is not particularly limited, but is preferably 20 to 100 mol%, particularly preferably 50 to 95 mol% of the whole polyvinyl acetate. When the degree of saponification of the polyvinyl acetate is within the above range, the effect of reducing the amount of adsorption of the biological substance is particularly excellent.

As the water-soluble resin having the first functional group in the side chain, for example, a resin represented by the following formula (Ia ) is preferable. Thereby, a uniform film can be formed at a practical wavelength of 300 to 500 nm, and the effect of reducing the adsorption of a biological substance can be particularly improved.

R represented by the formula (Ia ) of the water-soluble resin is not particularly limited as long as it is an alkyl group having a carbonyl and an amine, but for example, those represented by the following formula (III) are preferable.
Thereby, the synthesis of the first functional group can be easily performed.

The water-soluble resin having the first functional group in the side chain represented by the formula (Ia ) has a modification rate of the first functional group with respect to the water-soluble resin having the first functional group in the side chain, Although not particularly limited, 0.1 to 20 mol% is preferable, and 0.2 to 10 mol% is particularly preferable. If the denaturation rate is less than the lower limit, the water-soluble resin may not be sufficiently fixed to the biochemical danger surface, and if it exceeds the upper limit, the adsorption of biological substances is reduced. Effect may be reduced.

For the same reason, the degree of acetalization of the water-soluble resin having the first functional group in the side chain represented by the formula (Ia ) is preferably from 0.1 to 20 mol%, particularly from 0.2 to 10 mol% is preferable.
The water-soluble resin having the first functional group in the side chain may be in an uncured state, a cured state, or a semi-cured state between the uncured state and the cured state. It is preferable that Thereby, the eluate at the time of adding a sample can be reduced, and a surface having resistance to physical stimulation can be obtained.

  The water-soluble resin having the first functional group in the side chain is preferably formed on the surface of a biochemical instrument to be described later, and the thickness thereof is not particularly limited, but is preferably 2 to 3,000 nm, particularly 5 ˜2,000 nm is preferred. If the thickness is less than the lower limit, it may be difficult to form a uniform film. If the thickness exceeds the upper limit, a biological substance is taken into the water-soluble resin having the first functional group in the side chain. As a result, the effect of reducing the adsorption of the biological substance may be reduced.

Next, a biochemical instrument will be described.
The biochemical instrument is made of the resin material. In addition to making the biochemical instrument a disposable type, the resin material can be easily molded into various shapes.
Examples of the resin material include polypropylene resins, polyethylene resins, polyolefin resins such as ethylene-propylene copolymers, polystyrene resins such as polystyrene and acrylonitrile-butadiene-styrene resins, polycarbonate resins, polyethylene terephthalate resins, and polymethyl methacrylate. Methacrylic resin such as resin, vinyl chloride resin, polybutylene terephthalate resin, polyarylate resin, polysulfone resin, polyethersulfone resin, polyetheretherketone resin, polyetherimide resin, fluorine resin such as polytetrafluoroethylene, Examples thereof include acrylic resins such as polymethylpentene resin and polyacrylonitrile, and fiber-based resins such as propionate resin. Among these, a polypropylene resin is particularly preferable in terms of moldability, transparency, resistance to chemical and physical irritation required for biochemical instruments.

The weight average molecular weight of the resin material is not particularly limited, but is preferably 10,000 to 500,000, particularly preferably 20,000 to 100,000. When the weight average molecular weight is within the above range, the moldability of the biochemical instrument is excellent, and the molded product is excellent in water resistance.
The weight average molecular weight is, for example, G.M. P. C. It can obtain | require in conversion of styrene using.

  For the purpose of improving moldability and weather resistance, additives such as hydrocarbon-based, fatty acid amide-based lubricants, phenol-based and amine-based antioxidants are added to the resin material as long as the object of the present invention is not impaired. can do.

When the biochemical instrument is manufactured from the resin material, the biochemical instrument can be manufactured by, for example, injection molding, blow molding, or injection blow molding.
Examples of the biochemical instrument include liquid manipulation instruments such as pipettes and dispenser chips used for biochemical research, and containers such as centrifuge tubes, cryopreservation tubes, tubes, and multiwell plates. Among these, the contact time with a biological substance is relatively long, and it is used as tubes (specifically, blood collection containers or test tubes and sample containers used for protein analysis) used for sample preparation in various analyses. It is preferred that This improves the accuracy and sensitivity of the analysis results.

  The surface of the biochemical instrument is not particularly limited, but a second functional group is preferably formed. Thereby, fixation with the said 1st functional group can be strengthened, and the said water-soluble resin can be stably fixed to the surface of a biochemical instrument by it.

  Examples of the second functional group include a hydroxyl group, a carboxyl group, a carbonyl group, an amino group, and an amide group. Among these, at least one selected from a hydroxyl group, a carboxyl group, and a carbonyl group is preferable. Thereby, especially fixation with the 1st functional group can be strengthened.

The second functional group is preferably formed on the surface of the biochemical instrument, and the amount of the second functional group is not particularly limited, but is preferably 0.01 to 100 nmol / cm ² , particularly 0.05-10 nmol / cm < ² > is preferable. When the amount of the functional group is less than the lower limit value, the first functional group may not be sufficiently fixed, and when the upper limit value is exceeded, the characteristics of the base material itself change, as a biochemical instrument. May fail to meet the required performance.

  Examples of the method for forming the second functional group include a method for plasma treatment of the surface of the biochemical instrument, a method for irradiating with radiation, a method for graft polymerization, and the like. Among these, it is preferable to perform a plasma treatment method, particularly an oxygen plasma treatment. Thereby, the second functional group can be efficiently formed on the surface of the biochemical instrument.

As a method for fixing the water-soluble resin to the biochemical instrument, a method in which the first functional group and the surface of the biochemical instrument are covalently bonded, a hydrogen bond method, an ionic bond method, S- Examples include S-bonding methods. Among these, a covalent bond method is preferable. Thereby, the tolerance with respect to the chemical and physical irritation | stimulation of a surface can be improved. More specifically, it is preferable that the first functional group and the second functional group are covalently bonded. Thereby, the water-soluble resin having the first functional group in the side chain is efficiently fixed, and the resistance to chemical and physical stimulation on the surface can be further improved.
In addition to the covalent bond between the first functional group and the second functional group, the first functional group and a carbon-hydrogen bond formed on the surface of the biochemical instrument, carbon- A covalent bond with a carbon bond or the like may be mixed.

The adsorption rate [Q1] of the biological material of the biochemical instrument is not particularly limited, but is preferably 20% or less, particularly preferably 0.0001 to 10%, and most preferably 0.0002 to 5%. . When the adsorption amount is within the above range, it can be suitably used for a specimen storage device for clinical examinations that are required to have a particularly low adsorption rate of a biological substance. Furthermore, it can be suitably used as a biochemical instrument used for testing with a very small amount of sample.
The adsorption rate [Q1] of the biological substance can be determined by the following formula (1).
Adsorption rate (%) = Adsorption amount / Total amount of protein in solution (1)
For example, the adsorption rate of phosphate ( ¹²⁵ I) -labeled bovine serum immunoglobulin G at a concentration of 1.0E-6 g / mL, 1.0E-7 g / mL, 1.0E-8 g / mL, phosphate buffer (pH 7 4), dispense into the container of the present invention, and allow to stand at 37 ° C. for 1 hour. Thereafter, the washing is repeated 3 times with a phosphate buffer containing 0.05% by volume of Tween 20, and the container body is measured with a γ-ray counter.
The weight of iodine ( ¹²⁵ I) -labeled bovine serum immunoglobulin G remaining in the container body is determined from a separately prepared calibration curve, and the adsorption rate at each solution concentration is calculated.

The adsorption rate [Q2] of the biological substance after contacting the biochemical instrument of the present invention with a 50 volume% aqueous solution of acetonitrile for 10 minutes is not particularly limited, but is preferably 25% or less, particularly 5 to 5. 20% is preferred. In order for the adsorption rate [Q2] of the biological substance to be less than the lower limit value, it is necessary to limit the type of the biological substance, and if it exceeds the upper limit value, as a biochemical instrument used for testing with a very small amount of sample Cannot be suitably used.
Specifically, the adsorption rate [Q2] of the biological substance after contacting with a 50% by volume aqueous solution of acetonitrile for 10 minutes can be obtained as follows.
A 50 volume% aqueous solution of acetonitrile is dispensed into the container of the present invention, left to stand for 10 minutes, washed with pure water, dried, and the adsorption rate is calculated using the above formula (1).

  Examples of the biological substance include natural or synthetic peptides, proteins, enzymes, saccharides, lectins, viruses, bacteria, nucleic acids, DNA, RNA, antigens (for example, including recombinant antigens) antibodies, IgG. IgM. Immunoglobulins such as IgE: complement, CRP. Plasma proteins such as ferritin, α1 microglobulin, β2 microglobulin and their antibodies: tumor markers such as α-fetoprotein, carcinoembryonic antigen (CEA), prostatic acid phosphatase (PAP), CA19-9, CA-125 and Their antibodies: hormones such as luteinizing hormone (LH), follicle stimulating hormone (FSH), human ciliary gonadotropin (hCG), estrogen, insulin and their antibodies: HBV-related antigens (HBs.HBe.HBc). HIV. Substances related to virus infection such as ATL and their antibodies: bacteria such as diphtheria, botulinum, mycoplasma, syphilis treponema and their antibodies: protozoa such as toxoplasma, trichomonas, leishmania, tribanosome, malaria parasite and their antibodies: Drugs such as antiepileptic drugs such as phenytoin and phenobarbital, cardiovascular drugs such as quinidine and digoxinine, antiasthma drugs such as theophylline, antibiotics such as chloramphenicol and gentamicin and their antibodies: other enzymes, bacterial cells Examples include exotoxins (such as styrelidin O) and antibodies thereof. Among these, serum albumin or serum immunoglobulin G is preferable, and can be used for a wide range of studies including clinical diagnosis.

EXAMPLES Hereinafter, although this invention is demonstrated in detail based on an Example and a comparative example, this invention is not limited to this.
Example 1
Polypropylene resin (manufactured by Sumitomo Noblen Co., WP708) is used as a resin material, and injection molding (molding machine: manufactured by Nissei Plastic Industries 60t, cylinder temperature: 225 ° C-220 ° C-195 ° C-185 ° C, injection rate: 25%- 20% -15%, injection pressure: 40% -30% -25%, mold cooling: 30 ° C.). The obtained tube was subjected to plasma treatment (oxygen plasma for 5 minutes) using a plasma treatment apparatus (SERIES7000 manufactured by BRANSON / IPC) to form second functional groups on the surface of the tube.
In addition, the shape of the obtained tube was a V bottom tube having a height of 39 mm, an outer diameter of 10 mm, and a capacity of 1.5 mL.

Next, polyvinyl alcohol having an azide group in the side chain as a water-soluble resin having a first functional group in the side chain (AWP manufactured by Toyo Gosei Kogyo Co., Ltd., average polymerization degree of water-soluble resin 1,800, first functional group Was dissolved in pure water in a glass container light-shielded with an aluminum foil to prepare a 1.0 wt% solution.
The tube on which the second functional group is formed is dipped in a glass container shielded with aluminum foil for 1 minute, then taken out, dried at 40 ° C. for 60 minutes, and then subjected to UV light with a UV lamp at 0.1 mW. Irradiated with / cm ² × 5 minutes to cure the water-soluble resin having the first functional group on the side chain and then washed with pure water to obtain the biochemical instrument (tube) of the present invention.
On the surface of the obtained tube, a layer formed of a water-soluble resin having the first functional group in the side chain was formed with a thickness of 300 nm.

(Example 2)
The procedure was the same as Example 1 except that the concentration of the pure aqueous solution of the water-soluble resin having the first functional group in the side chain was changed as follows.
A glass container in which polyvinyl alcohol having an azide group in the side chain (AWP manufactured by Toyo Gosei Kogyo Co., Ltd., average polymerization degree of water-soluble resin 1,800, first functional group modification rate 0.6 mol%) is shielded with aluminum foil The solution was dissolved in pure water to prepare a 3.0 wt% solution.
On the surface of the obtained tube, a layer formed of a water-soluble resin having the first functional group in the side chain was formed with a thickness of 1,100 nm.

(Example 3)
The same procedure as in Example 1 was performed except that the concentration of the pure aqueous solution of the water-soluble resin having the first functional group in the side chain was changed as follows.
A glass container in which polyvinyl alcohol having an azide group in the side chain (AWP manufactured by Toyo Gosei Kogyo Co., Ltd., average polymerization degree of water-soluble resin 1,800, first functional group modification rate 0.6 mol%) is shielded with aluminum foil The solution was dissolved in pure water to prepare a 0.001% by weight solution.
On the surface of the obtained tube, a layer formed of a water-soluble resin having the first functional group in the side chain was formed with a thickness of 4 nm.

(Example 4)
The same procedure as in Example 1 was performed except that the tube was not previously plasma-treated and the second functional group was not formed.
On the surface of the obtained tube, a layer formed of a water-soluble resin having the first functional group in the side chain was formed with a thickness of 200 nm.

(Example 5)
The same procedure as in Example 1 was carried out except that UV light was not irradiated by a UV lamp, pure water was not washed without sufficiently curing the water-soluble resin having the first functional group in the side chain.

(Example 6)
Methyl pentene (TPX) resin (manufactured by Mitsui Petrochemical Co., Ltd., RT-31) was used as the resin material, and the same procedure as in Example 1 was performed except that the injection molding conditions were as follows.
Injection molding machine: Nissei Plastic Industries 60t, cylinder temperature: 300 ° C-280 ° C-265 ° C-255 ° C, injection speed: 40% -30% -10%, injection pressure: 60% -30% -25% Mold cooling: performed at 50 ° C.
On the surface of the obtained tube, a layer formed of a water-soluble resin having the first functional group in the side chain was formed with a thickness of 250 nm.

(Comparative Example 1)
Polyvinyl alcohol having no functional group in the side chain as a water-soluble resin: Examples except that an average polymerization degree of about 1,500 and a saponification degree of 86 to 90 mol% (manufactured by Wako Pure Chemical Industries, Ltd., 160-03055) were used Same as 1.

The following evaluation was performed about the obtained container. The evaluation items are shown together with the contents. The obtained results are shown in Table 1.
1. Comparison of Adsorbability of Biological Substances 1.1 Bovine Serum Immunoglobulin Adsorbability The adsorptivity of bovine serum immunoglobulin was evaluated as follows. Bovine serum immunoglobulin (manufactured by Bovine gamma globulin standard 23210 PIERCE) was labeled with iodine ( ¹²⁵ I) and diluted with a phosphate buffer (pH 7.4) to a concentration of 1.0E ^-7 g / mL. It dispensed into the container obtained by the comparative example, and left still at 37 degreeC for 1 hour. Thereafter, washing was repeated 3 times with 0.05 vol% Tween20-containing phosphate buffer, and the container body was measured with a γ-ray counter.
The weight of iodine ( ¹²⁵ I) -labeled bovine serum immunoglobulin remaining in the container body was determined from a separately prepared calibration curve, and the adsorption rate at each solution concentration was calculated.

1.2 Bovine serum albumin adsorptivity The adsorptivity of bovine serum albumin was evaluated as follows. A solution obtained by diluting bovine serum albumin (manufactured by 23209 PIERCE) to 0.5 μg / mL was dispensed at 1.0 mL each into the containers obtained in each Example and Comparative Example, and incubated at 37 ° C. for 1 hour. Washed 3 times with a phosphate buffer pH 7.4 containing 0.05% by volume tween20.
Next, as a blocking, a phosphate buffer solution pH 7.4 containing 3.0% by weight skimmed milk was dispensed in 1.5 mL portions, incubated at 37 ° C. for 1 hour, and then phosphate buffer solution pH 7.4 containing 0.05% by volume tween20. And washed 3 times. Next, a solution obtained by diluting peroxidase-labeled anti-bovine serum albumin antibody (55285 CAPPEL) to 1.0 μg / mL with phosphate buffer pH 7.4 was dispensed at 1.0 mL in each container and incubated at room temperature for 30 minutes. After washing with 0.05 volume% tween 20 phosphate buffer pH 7.4 three times, color was developed using a peroxidase color development kit (SUMILON ML-1120T manufactured by Sumitomo Bakelite Co., Ltd.) and a plate reader was used. The absorbance at 450/630 nm was measured.
The weight of peroxidase-labeled anti-bovine serum albumin antibody = bovine serum albumin remaining in the container body was determined from a separately prepared calibration curve, and the adsorption rate was calculated.

1.3 Adsorbability of peroxidase-labeled avidin The adsorptivity of peroxidase-labeled avidin was evaluated as follows. A solution obtained by diluting peroxidase-labeled avidin (43-4423, manufactured by ZYMED) to 0.5 μg / mL was dispensed into each of the containers obtained in each Example and Comparative Example in an amount of 1.0 mL and allowed to stand at room temperature for 1 hour. Then, the plate was washed three times with a phosphate buffer solution pH 7.4 containing 0.05% tween 20, and developed using a peroxidase coloring kit (SUMILON ML-1120T manufactured by Sumitomo Bakelite Co., Ltd.), and then a plate reader was used. The absorbance at 450/630 nm was measured.
The weight of peroxidase-labeled avidin remaining in the container body was determined from a separately prepared calibration curve, and the adsorption rate was calculated.

2. Measurement of eluate The eluate was measured with a UV spectrophotometer.
Pure water was dispensed in 1.5 mL increments into the containers obtained in each Example and Comparative Example, and the dispensed pure water was recovered after incubation at 37 ° C. for 24 hours, and the UV spectrophotometer measured 220 nm to 300 nm. The presence or absence of absorption was confirmed.

3. Solvent resistance The solvent resistance was evaluated by the adsorption rate of bovine serum immunoglobulin G after the following treatment.
A 50 volume% aqueous solution of acetonitrile was dispensed in 1.5 mL aliquots into the containers obtained in each of the examples and comparative examples, left standing for 10 minutes, washed with pure water, and evaluated as described above (bovine serum immunoglobulin adsorptivity). Thus, the adsorptivity of bovine serum immunoglobulin G was evaluated.

4). Centrifugal strength Centrifugal strength was evaluated using a centrifuge.
Dispense 1.5 mL of pure water into the containers obtained in each Example and Comparative Example, and set in a centrifuge (CF16RX type Hitachi Multi-purpose Small Centrifuge Rotor # 11). After centrifuging for minutes, the container was visually checked for deformation and cracking.

5. Heat resistance The heat resistance was evaluated by the deformation of the container after the following treatment and the adsorption rate of bovine serum immunoglobulin G.
Dispense 1.5 mL of pure water into the containers obtained in each Example and Comparative Example, leave it in a thermostatic bath at 95 ° C. for 10 minutes, discard the pure water, and visually check the container for deformation and cracking. did.
Further, the adsorptivity of bovine serum immunoglobulin G was evaluated by the method for evaluating the adsorptivity of bovine serum immunoglobulin.

6). Evaluation of cold resistance Cold resistance was evaluated by the deformation of the container after the following treatment and the adsorption rate of bovine serum immunoglobulin G.
Dispense 1.5 mL of pure water into the containers obtained in each Example and Comparative Example, and leave it in a deep freezer at −80 ° C. for 24 hours, then discard the pure water, and visually observe deformation and cracking of the tube body. It was confirmed.
Further, the adsorptivity of bovine serum immunoglobulin G was evaluated by the method for evaluating the adsorptivity of bovine serum immunoglobulin.

As is apparent from Table 1, Examples 1 to 6 showed that the amount of adsorption of the biological substance could be reduced. Thereby, it can use suitably for a blood collection container or a test tube and a sample container used for protein analysis.
In addition, in Examples 1 to 3 and 6, the amount of the eluate was particularly small, thereby preventing the direct influence on the sample and the influence on the measurement result due to the dissolution of the water-soluble resin. it can.
Examples 1 to 6 were also excellent in solvent resistance and centrifugal strength.
Moreover, Examples 1-6 were excellent also in heat resistance, and Examples 1-3 and 6 also had especially few adsorption amount of the biological substance after heat processing.
Moreover, Examples 1-6 were excellent also in cold resistance, and Example 1, 2, 4, and 6 also had especially few adsorption amount of the biological substance after freezing process.
As described above, Examples 1 to 6 have been shown to be suitable for a container for analyzing a low-concentration biological substance because the adsorption rate of the biological substance is low.

  The biochemical instrument of the present invention can be suitably used for biochemical research such as genetic engineering, protein engineering, and clinical laboratory science. There is no loss due to adsorption of samples such as biological substances, organic solvents can be used, and highly accurate and sensitive analysis can be performed. Specifically, it can be used as a pipette, a dispenser chip, a storage container, a dilution container, and the like, which are instruments particularly used for analyzing a biological substance.

Claims (7)

A water-soluble resin having a first functional group on a side chain is fixed to a biochemical instrument composed of a resin material,
The water-soluble resin having the first functional group in the side chain is polyvinyl alcohol obtained by saponifying polyvinyl acetate, and the polyvinyl alcohol further includes a structural unit represented by the following formula (Ia ). Biochemical instrument characterized by being a thing.

The biochemical instrument according to claim 1, wherein a second functional group is formed on a surface of the biochemical instrument. The biochemistry according to claim 2, wherein the water-soluble resin is fixed to the biochemical instrument mainly by covalently bonding the first functional group and the second functional group. Appliances. The biochemical instrument according to any one of claims 1 to 3, wherein an adsorption rate of the biological substance is 20% or less. The biochemical instrument according to any one of claims 1 to 4, wherein an adsorption rate of a biological substance after contact with a 50 vol% aqueous solution of acetonitrile for 10 minutes is 25% or less. The biochemical instrument according to claim 4 or 5, wherein the biological substance is bovine serum albumin or bovine serum immunoglobulin G. The biochemical instrument according to any one of claims 1 to 6, wherein the biochemical instrument is used as a tube.