JPH08259636A - Cationic polymer electrolyte copolymer latex - Google Patents

Abstract

PURPOSE: To obtain an electrolyte copolymer latex free from interference problem in the use as an antibody-adsorbing latex diagnostic and useful as an immunoassay material absolutely without using a surfactant by grafting an olefin monomer to a cationic derivative of a water-soluble linear polysaccharide in water. CONSTITUTION: This latex is composed of a copolymer of formula IV produced by grafting (A) a cationic derivative of a water-soluble linear polysaccharide expressed by formula I [X is (CH2 )n R1 (R1 is NH<+> 3 , NH<+> (CH3 )2 , etc.; (n) is 1-3), etc.; 0<a<3; 50,0O0>=x>=5] with (B) an olefin compound of formula II [R4 to R6 are each H or CH3 ; R7 is formula III (R8 is H, a 1-12C alkyl, etc.), etc.; (m) is 20-200,000] in water. The copolymer contains the component A as the stem polymer and the component B as the graft chain bonded to the stem polymer by abstracting the H atom of the OH group of the polymer. The graft ratio of the copolymer is 2-5,000%.

Description

Detailed Description of the Invention

[0001]

INDUSTRIAL APPLICABILITY The cationic copolymer of water-soluble linear polysaccharide-olefin monomer graft copolymer of the present invention is a cationic derivative of water-soluble linear polysaccharide having a hydroxyl group, and is an olefin monomer in water. Graft-polymerize
It is intended to produce a latex polymerization product useful as an immunoassay material. INDUSTRIAL APPLICABILITY The present invention suggests that a cationic derivative of a linear polysaccharide having a hydroxyl group and a water-soluble olefin monomer can be generally graft-polymerized in water to produce a latex polymerization product useful as an immunoassay material. It is a thing.

[0002]

2. Description of the Related Art Conventionally, most methods for producing latex polymerization products as immunoassay materials are those obtained by emulsion polymerization in an aqueous solution in the presence of a surfactant. Things are desired. This is because the surfactant present in the aqueous solution affects the action as a latex diagnostic agent.

[0003]

The cationic derivative of a water-soluble linear polysaccharide-olefin monomer graft copolymer of the present invention is a cationic derivative of a water-soluble linear polysaccharide having a hydroxyl group, and an olefin in water. Graft polymerization of a monomer by a suspension polymerization method or an emulsion polymerization method to produce a latex polymerization product useful as an immunoassay material, and a cationic derivative of the water-soluble linear polysaccharide of the present invention-monomer olefin The polymer graft copolymer latex is used for the antibody-adsorbed latex diagnostic agent according to the claims, which is a latex diagnostic agent. Emulsion polymerization is a polymerization method in which an olefin monomer is suspended in an aqueous solution and usually emulsified by using a surfactant or the like. Specifically, it is a hydrogen bond with a monomer or a growing chain, a Coulomb force, a charge. It is a polymerization method in which a polymer chain is polymerized and grown by interaction at a water-solvent interface by transfer interaction, van der Waals force, etc. to form fine particles in an aqueous solution. Usually, the polymerization product is present as a mixture of polymerized monomer and surfactant.
This surfactant, which is considered as an impurity, may interfere when it is used in the antibody-adsorbed latex diagnostic agent described in the claims which is a latex diagnostic agent, which is a problem.

[0004]

INDUSTRIAL APPLICABILITY The present invention is a product obtained by graft-polymerizing an olefin monomer in water with a cationic derivative of a water-soluble linear polysaccharide having a hydroxyl group. The structures of the growing olefin monomer growth chain and the resulting copolymer chain are described as chemical structural formulas (2) and (3), respectively. Each bond is shared by chain transfer of olefin monomer double bond resulting from radical generation by abstraction of hydrogen atom of hydroxyl group (as proton by oxidation of initiator) of cationic derivative of water-soluble linear polysaccharide. It is clear that it is a bond. This is carried out by using tetravalent cerium ion as an initiator, and since no surfactant is used, it is very useful because it is not disturbed when it is used as an antibody-adsorbed latex diagnostic agent. The cationic derivative of water-soluble linear polysaccharide-olefin monomer graft copolymer latex of the present invention is intended to be used as an antibody-adsorbed latex diagnostic agent according to the claims, which is a latex diagnostic agent. Is clear from the claim format. That is, the olefin monomer is graft-polymerized in water to a water-soluble polymer having a hydroxyl group,
The production of latex polymerization products useful as immunoassay diagnostic materials is a major part of the essential part of the constitution of the present invention, and olefin monomer graft copolymers such as polysaccharides can also achieve the same purpose. . The so-called graft-free graft copolymer that grows at the interface between the solvent and the solute is a useful substance having a wide variety of uses. In particular, attention has been paid to filtration membranes and biomaterials in addition to immunoassay materials. Focusing on its hydrophilicity, it can be applied to artificial kidney membranes, components, blood substitutes, and contact lenses.

[0005]

Detailed Description of the Structure of the Invention Hereinafter, the cationic derivative of linear polysaccharide-olefin monomer graft copolymer of the present invention will be described in detail. The cationic derivative of linear polysaccharide-olefin monomer graft copolymer is obtained through the following two steps. ◆. Preparation of Cationic Derivative of Linear Polysaccharide The unit formula of the cationic derivative of linear polysaccharide is [C
6 H7 O2 (OH) 3-a. (OX) a] x
* H2O (1) [In formula, X is-(CH2) nR1 (R1 is -NH.
2, -N (CH3) 2, -N (C2H5) 2
, -N + (C2H5) 3, -N + (C2
H5) 2 (C2H5) N (C2H5) 2
, -C6 H4.NH2, -CO.C6 H4.
A group selected from the group consisting of NH2, n = 1 to an integer of 1 to 3, or -COR2 (R2 is -CH2.NH2.
Or -C6H4.NH2) or -CH2CH
(OH) .CH2 R3 (R3 is -NH2, -N
(CH3) 2, -N (C2H5) 2, -N +
A group selected from the group consisting of (C2H5) 3),
Alternatively, -NH.CH2.CH2, a is a positive number of 0 <a <3, and x is an integer of 50,000 ≧ x ≧ 5]. In the above (1), the hydroxyl group of the cationic derivative of the linear polysaccharide is partially substituted with an acid group such as carboxymethyl group or sulfate group by an ether bond, or partially substituted with an alkyl group. It may have a cationic functional group represented by X in the formula. Usually, the cationic derivatives of these linear polysaccharides are obtained by the Schotten-Baumann reaction in an alkaline solution of the hydroxyl group of the linear polysaccharide and the chlorine compound of the above cationic substituent represented by XCl. The linear polysaccharides referred to here include those that can be industrially produced by fermentation such as dextran and pullulan. As the olefin monomer to be graft-polymerized, one having a general formula represented by the following formula can be considered. [Wherein R5, R6 and R7 are each selected from a hydrogen atom or CH3, Group, cyclohexyl group, C1-C4 hydroxyalkyl group, C1-C8 aminoalkyl group, C1
To C8 dialkylaminoalkyl group, glycidyl group, tetrahydrofuran group, C1 to C4 lower alkyl substituted tetrahydrofuran group, benzyl group and (-C
H2CH2-O-) yCH2CH2OH group (where y is a positive number of 1 to 10), -N (R10) 2 (R10
Is a hydrogen atom or a C1-C4 alkyl group, two R
10 may be the same or different), (R11 represents a C1-C8 alkyl group), a phenyl group, a pyridine group, a tolyl group, and a pyrrolidone group]. Specifically, α, β-unsaturated acid alkyl esters such as acrylic acid and methacrylic acid, lower alkyl-substituted cyclohexyl esters such as cyclohexyl ester, 2-hydroxyethyl ester, 2-hydroxypropyl ester, and 2-hydroxy. Butyl ester; acrylamide, methacrylamide, acrylamide, acryl- or methacryl-dimethylamide, C1 to C3 aminoalkyl ester of the above α, β-unsaturated acid, C1 to C3 dialkylaminoalkyl ester, glycidyl ester, tetrahydrofurfuryl Esters, benzyl esters, polyethylene glycol monoesters; α, β-unsaturated acid nitrile groups such as acrylonitrile and methacrylonitrile; vinyl alcohol, methyl vinyl alcohol Vinyl alcohol, vinyl acetate such as vinyl acetate, vinyl propionate, and vinyl butyrate, and its C1 to C3 alkyl esters of methyl-substituted vinyl alcohol;
Possible examples include styrene, vinyltoluene; vinylpyrrolidone; vinylmethylpyrrolidone. ◆. The adjustment reaction of the graft copolymer is usually performed in an aqueous solution. That is, the above olefin monomer is added to an aqueous solution of a cationic derivative of linear polysaccharide and an initiator is added to react. As the initiator, a tetravalent cerium salt, a tetravalent manganese salt, a ferric salt-hydrogen peroxide is usually used, but potassium persulfate (K
Radical initiators such as PS), azobisisobutylnitrile (AIBN) and benzoyl peroxide (BPO) are also used. The reaction temperature can be widely selected from normal temperature to 80 ° C.
If necessary, the reaction is continued by replacing with nitrogen.
Each bond is a covalent bond due to chain transfer of an olefin monomer double bond by radical generation due to proton abstraction of a cationic derivative of water-soluble linear polysaccharide. In this reaction, the product occurs in latex. This latex polymer is generally water, alcohol
Or insoluble in organic solvents such as acetone and tetrahydrofuran, but can be easily formed into a film by the casting method or the like. Alternatively, an insoluble solvent such as alcohol is added in excess to obtain a precipitate, and then a molded product can be easily produced by a hot pressing method or the like. For the above purpose, the ratio of the trunk polymer to the graft polymer or the polymerization degree ratio thereof in the graft polymer can be variously selected according to the purpose. In the graft polymerization, the polymerization rate is determined by the graft rate (%).
This is defined by the following formula: graft ratio (%) = (amount of graft-polymerized monomer / amount of trunk polymer in graft copolymer) × 100. In the present invention, the olefin compound is formed as a graft chain, and a graft ratio in the range of 2% to 5000% is considered appropriate. It has been repeatedly stated that the present invention is a product obtained by graft-polymerizing an olefin monomer underwater into a cationic derivative of a water-soluble linear polysaccharide having a hydroxyl group, but the structure of the resulting copolymer chain is claimed. As represented by the chemical structural formula in the range of, the formula (1) and the formula (2) are represented by the following formula (3) [[C6H7O2 (OH) 3-a. (OX) a]. x.H2O] [R4 CR5 R6 CR7] m (3) Each of the bonding relationships is a chain of olefin monomer double bonds due to radical generation by abstraction of proton of hydroxyl group of cationic derivative of water-soluble linear polysaccharide. It is a covalent bond due to movement.

[0006]

【Example】

Example 1 2 g of DEAE (diethylaminoethyl) dextran hydrochloride having a nitrogen content of 5% and having dextran having an average molecular weight Mw of 500,000 as a matrix was dissolved in 50 ml of water, and then methanol was added.
5 ml and 30 ml of methyl methacrylate (MMA) were added, the air in the reaction solution and the reaction vessel was sufficiently replaced with nitrogen gas, and then the mixture was stirred well and cerium ammonium nitrate nitric acid nitrate dissolved in 15 ml of 0.1N nitric acid. 190
Add mg to start the reaction. The reaction is carried out at 30 ° C. for 1 hour to form a latex. At the end of the reaction, 3 ml of a 1% solution of hydroquinone was used as a terminator. Then, the reaction solution was poured into 3-fold amount of methanol to obtain a precipitate. The precipitate was thoroughly washed with hot water, centrifuged, dried under reduced pressure at 50 ° C., and the dried product was placed in a Soxhlet extractor and subjected to acetone extraction for 24 hours to obtain a DEAE (diethylaminoethyl) dextran-MMA copolymer. 1.5 g of hydrochloride was obtained. Nitrogen content 2.0% Graft ratio 25% vs. DEAE-dextran yield 30% This is insoluble in water, which is a good solvent for DEAE dextran hydrochloride, and acetone, which is a good solvent for polymethylmethacrylate. When the infrared absorption spectrum of this product is examined, absorption of a carbonyl group which is not found in DEAE-dextran hydrochloride is found near 1730 cm −1. Example 2 After the same reaction as in Example 1, the reaction-completed solution of the latex was not poured into methanol, but the mixture was subjected to a water-falling test to remove unreacted materials and an initiator, and then the homopolymer A mixed DEAE dextran-MMA copolymer latex was obtained. This is useful as a reagent for detecting rheumatoid factor and shows the test results. Test Method An equal amount of antibody in a 2% solution of this latex [ie 2
Glycine-Nac so as to become 0 μg / mg latex
l buffer-γ globulin (GB) (0.05 mol /
1, diluted with PH 8.2)] while stirring 1
Add dropwise and stir at room temperature for 2 hours, then (400
Centrifuge at 0 rpm for 30 minutes and repeat resuspension with GB)
Finally at GB with 0.5% BSA (bovine albumin) 1
Store at 4 ° C as a% suspension. A drop of the sensitized latex liquid of the present invention was dropped on a wiped glass plate with a capillary tube,
Rheumatoid factor-positive serum diluted 1/20 with GB in advance was added dropwise to a 0.05 ml latex solution on a glass plate with a capillary tube, stirred and shaken. When judged, aggregates were found. When the same operation was performed with a known 1/20 diluted rheumatoid factor negative serum, no aggregate was observed. Example 3 4 g of DEAE (diethylaminoethyl) -pullulan hydrochloride having a nitrogen content of 4% and having pullulan having an average molecular weight Mw of 200,000 as a matrix was dissolved in 80 ml of water, and then methanol 10 was added.
ml, 35 ml of styrene monomer were added, the reaction solution and the air in the reaction vessel were sufficiently replaced with nitrogen gas, and then 200 mg of ceric ammonium nitrate nitrate dissolved in 30 ml of 0.1N nitric acid was added with thorough stirring. Start the reaction. The reaction is carried out at room temperature for 1 hour to form a latex. At the end of the reaction, 3 ml of a 1% solution of hydroquinone was used as a terminator. The subsequent purification and drying steps were performed in the same manner as in Example 1 to obtain 6 g of DEAE (diethylaminoethyl) pullulan-styrene copolymer hydrochloride. Nitrogen content 1.14% Grafting ratio 350% DEAE pullulan yield 43% Example 4 After carrying out the same reaction as in Example 3, the reaction completed solution of the latex was not poured into methanol but permeated in water. After folding, the unreacted material and the initiator were removed to obtain a DEAE pullulan-styrene copolymer latex in which a homopolymer was mixed. This was useful as a reagent for detecting rheumatoid factor. As in Example 2, a 2% solution of this latex in an equal amount of the antibody, [ie glycine-Nacl buffer-γ globulin (GB) (0.05 mol / l, PH 8.2) to give a 20 μg / mg latex).
The latex solution diluted with and sensitized with a rheumatoid factor-positive serum was visually judged by the procedure of Example 2 for the presence or absence of agglutination, but agglutinates were observed in the rheumatoid factor-negative serum. There wasn't. Example 5 4 g of AE (aminoethyl) dextran hydrochloride having a nitrogen content of 5% and having dextran having an average molecular weight Mw of 40,000 as a matrix
Was dissolved in 90 ml of water, then 5 ml of methanol and 20 ml of butyl methacrylate were added, and the reaction solution and air in the reaction vessel were sufficiently replaced with nitrogen gas, and then dissolved in 15 ml of 0.1N nitric acid while stirring well. 50 mg of ceric ammonium nitrate nitrate is added to start the reaction. The reaction is carried out for 30 minutes at room temperature to form a latex. At the end of the reaction, 3 ml of a 1% solution of hydroquinone was used as a terminator. Subsequent purification and drying steps were performed in the same manner as in Example 1 to obtain 6 g of hydrochloride of AE (aminoethyl) dextran-butyl methacrylate copolymer. Nitrogen content 2.0% Graft ratio 300% vs. AE-dextran yield 30% This is insoluble in water, which is a good solvent for AE dextran hydrochloride, and in acetone, which is a good solvent for polybutyl methacrylate. Example 6 After carrying out the same reaction as in Example 5, the reaction-completed solution of the latex was not poured into methanol, but the solution was subjected to a water-induced disintegration to remove unreacted substances and an initiator. A mixed AE (aminoethyl) dextran-butyl methacrylate copolymer latex was obtained. This was useful as a reagent for detecting rheumatoid factor. As in Example 2, a 2% solution of this latex in an equal amount of the antibody [ie glycine-to give 20 μg / mg latex-
Nacl buffer-γ globulin (GB) (0.05 m
The latex solution diluted with ol / l, PH 8.2) and sensitized was visually evaluated for the presence or absence of aggregation in the rheumatoid factor positive serum according to the procedure of Example 2. No aggregates were found in the negative serum. Example 7 Nitrogen content 3 with pullulan having an average molecular weight Mw of 30,000 as a matrix
% HPTMA (2-hydroxypropyltrimethylammonium) -pullulan hydrochloride 4 g was dissolved in 100 ml water, then 30 ml methyl acrylate monomer was added,
After sufficiently replacing the air in the reaction solution and the reaction vessel with nitrogen gas, while stirring well, 200 mg of ceric ammonium nitrate nitrate nitrate dissolved in 20 ml of 0.1N nitric acid
To start the reaction. The reaction is carried out at room temperature for 1 hour to form a latex. At the end of the reaction, 4 ml of a 1% hydroquinone solution was used as a terminator. Subsequent purification and drying steps were performed in the same manner as in Example 1 to obtain 2 g of hydrochloride of HPTMA (2-hydroxypropyltrimethylammonium) pullulan-methyl acrylate copolymer. Nitrogen content 2% Graft ratio 150% HPTMA pullulan yield 33% Example 8 After the same reaction as in Example 7, the reaction was conducted in water to remove unreacted materials and an initiator to obtain a homopolymer. A mixed HPTMA (2-hydroxypropyltrimethylammonium) pullulan-methyl acrylate copolymer latex was obtained. This was useful as a reagent for detecting rheumatoid factor. As in Example 2, a 2% solution of this latex in an equal amount of antibody [ie 20 μg / mg
Glycine-Nacl buffer to become latex
γ-globulin (GB) (0.05 mol / l, PH8.
The latex liquid diluted and sensitized in 2) was visually observed for the presence or absence of aggregates in the rheumatoid factor-positive serum according to the procedure of Example 2, but aggregates were observed in the rheumatoid factor-negative serum. I couldn't see it.

[Procedure amendment]

[Submission date] May 7, 1993

[Procedure Amendment 1]

[Document name to be amended] Statement

[Correction target item name] Full text

[Correction method] Change

[Correction content]

[Document name] Statement

Title: Cationic polyelectrolyte copolymer latex

[Claims]

Embedded image In, the formula of the unit of the cationic derivative of the water-soluble linear polysaccharide is

Embedded image The formula of the unit of the polymer of the olefin compound is represented by

Embedded image , A cationic derivative of a water-soluble linear polysaccharide is used as a trunk polymer, and its hydroxyl group is used as a binding site by abstracting a hydrogen atom, and an olefin compound is used as a graft chain. The graft ratio is in the range of 2% to 5000% ( Conversion
Water obtained by grafting an olefin monomer onto the cationic partially substituted polysaccharide of the water-soluble linear polysaccharide represented by the above (Chemical Formula 1), which is composed of 2) and this (Chemical Formula 3). A latex comprising a copolymer obtained by grafting an olefin monomer in water on a cationic partially substituted soluble linear polysaccharide.

In [Chemical Formula 1], cationic induction of water-soluble linear polysaccharides
The formula for the conductor unit is

Embedded image in which the polymer of the olefin compound is represented by
The formula for rank is

Embedded image is represented by, a water-soluble linear polysaccharide cationic derivative as a trunk polymer, the hydroxyl group as a binding site by a hydrogen atom abstraction, the olefinic compound is made as a graft chain, 5000% graft ratio of 2% (Chemical Formula 2) in the range of (Chemical Formula 2) and this (Chemical Formula 3), and is obtained by grafting an olefin monomer in water onto the cationic partially substituted form of the water-soluble linear polysaccharide represented by the above (Chemical Formula 1). A latex diagnostic agent obtained by adsorbing an antibody on a latex made of a copolymer.

Detailed Description of the Invention

[0001]

INDUSTRIAL APPLICABILITY The cationic copolymer of water-soluble linear polysaccharide-olefin monomer graft copolymer of the present invention is a cationic derivative of water-soluble linear polysaccharide having a hydroxyl group, and is an olefin monomer in water. Graft-polymerize
It is intended to produce a latex polymerization product useful as an immunoassay material. INDUSTRIAL APPLICABILITY The present invention suggests that a cationic derivative of a linear polysaccharide having a hydroxyl group and a water-soluble olefin monomer can be generally graft-polymerized in water to produce a latex polymerization product useful as an immunoassay material. It is a thing.

[0002]

2. Description of the Related Art Most conventional methods for producing latex polymerization products as immunoassay materials are those obtained by emulsion polymerization in an aqueous solution in the presence of a surfactant, and those of soapless without a surfactant. Is desired. This is because the surfactant present in the aqueous solution affects the action as a latex diagnostic agent.

[0003]

The cationic derivative of a water-soluble linear polysaccharide-olefin monomer graft copolymer of the present invention is a cationic derivative of a water-soluble linear polysaccharide having a hydroxyl group, and an olefin in water. Graft polymerization of a monomer by a suspension polymerization method or an emulsion polymerization method to produce a latex polymerization product useful as an immunoassay material, and a cationic derivative of the water-soluble linear polysaccharide of the present invention-monomer olefin The polymer graft copolymer latex is used for the antibody-adsorbed latex diagnostic agent according to the claims, which is a latex diagnostic agent. Emulsion polymerization is a polymerization method in which an olefin monomer is suspended in an aqueous solution and usually emulsified by using a surfactant, and more specifically, it is a hydrogen bond, Coulomb force, charge transfer interaction with a monomer or a growing chain. It is a polymerization method in which a polymer chain is polymerized and grown by interaction at a water-solvent interface by action, van der Waals force, etc., to form fine particles in an aqueous solution. Usually, the polymerization product is present as a mixture of polymerized monomer and surfactant.
This surfactant, which is considered as an impurity, may interfere when it is used in the antibody-adsorbed latex diagnostic agent described in the claims which is a latex diagnostic agent, which is a problem.

[0004]

INDUSTRIAL APPLICABILITY The present invention is a product obtained by graft-polymerizing an olefin monomer in water with a cationic derivative of a water-soluble linear polysaccharide having a hydroxyl group. The structures of the growing chain of the olefin monomer and the resulting copolymer chain are described as the chemical structural formulas ( Chemical Formula 3) and ( Chemical Formula 1) , respectively. Each bond is shared by chain transfer of olefin monomer double bond resulting from radical generation by abstraction of hydrogen atom of hydroxyl group (as proton by oxidation of initiator) of cationic derivative of water-soluble linear polysaccharide. It is clear that it is a bond. This is carried out by using tetravalent cerium ion as an initiator, and since no surfactant is used, it is very useful because it is not disturbed when it is used as an antibody-adsorbed latex diagnostic agent. The cationic derivative of water-soluble linear polysaccharide-olefin monomer graft copolymer latex of the present invention is intended to be used as an antibody-adsorbed latex diagnostic agent according to the claims, which is a latex diagnostic agent. It is clear from the claimed format that the water-soluble polymer having a hydroxyl group is graft-polymerized with an olefin monomer in water to produce a latex polymerization product useful as an immunoassay diagnostic material. It is an essential part of the constitution that is essential to the constitution of (1), and an olefin monomer graft copolymer such as a polysaccharide can also achieve the same purpose. Such a graft copolymer which grows at an interface between a solvent and a solute, which is called soap-free, is a useful substance having a wide variety of uses. In particular, in addition to immunoassay materials, they are attracting attention as membranes and biomaterials. Focusing on its hydrophilicity, it can be applied to artificial kidney membranes, components, blood vessels substitutes, and contact lenses.

[0005]

Detailed Description of the Structure of the Invention Hereinafter, the cationic derivative of linear polysaccharide-olefin monomer graft copolymer of the present invention will be described in detail. The cationic derivative of linear polysaccharide-olefin monomer graft copolymer is obtained through the following two steps. I. Preparation of Cationic Derivatives of Linear Polysaccharides The formula for the units of cationic derivatives of linear polysaccharides is

## STR2 ## In the above (Chemical formula 2) , the hydroxyl group of the cationic derivative of this linear polysaccharide is partially substituted with an acid group such as carboxymethyl group or sulfate group by an ether bond or partially substituted with an alkyl group. It may have a cationic functional group represented by X. Usually, the cationic derivatives of these linear polysaccharides are obtained by the Schotten-Baumann reaction in an alkaline solution of the hydroxyl group of the linear polysaccharide and the chlorine compound of the above cationic substituent represented by XCl. The linear polysaccharides referred to here include those that can be industrially produced by fermentation such as dextran and pullulan. As the olefin monomer to be graft-polymerized, one having a general formula represented by the following formula can be considered.

Embedded image Specifically, an alkyl ester of α, β-unsaturated acid such as acrylic acid or methacrylic acid, a lower alkyl-substituted cyclohexyl ester such as cyclohexyl ester, 2-hydroxyethyl ester, 2-hydroxypropyl ester. 2-hydroxybutyl ester; acrylamide, methacrylamide, acrylamide, acryl- or methacryl-dimethylamide,
The alpha, beta-amino alkyl esters of C ₁ -C ₃ unsaturated acids, dialkylaminoalkyl esters of C ₁ -C _3, glycidyl ester, tetrahydrofurfuryl ester, benzyl ester, polyethylene glycol monoesters, acrylonitrile, methacryl Nitrile groups of α, β-unsaturated acids such as ronitrile; vinyl alcohol, methyl vinyl alcohol, dimethyl vinyl alcohol; vinyl alcohol such as vinyl acetate, vinyl propionate and vinyl butyrate, and C ₁ -of methyl substituted vinyl alcohol thereof. C ₃ alkyl ester; styrene;
Possible examples are vinyltoluene; vinylpyrrolidone; vinylmethylpyrrolidone. II. The adjustment reaction of the graft copolymer is usually performed in an aqueous solution. That is, the above olefin monomer is added to an aqueous solution of a cationic derivative of linear polysaccharide and an initiator is added to react. As the initiator, a tetravalent cerium salt, a tetravalent manganese salt, a ferric salt-hydrogen peroxide is usually used, but potassium persulfate (K
Radical initiators such as PS), azobisisobutylnitrile (AIBN) and benzoyl peroxide (BPO) are also used. The reaction temperature can be widely selected from normal temperature to 80 ° C.
If necessary, the reaction is continued by replacing with nitrogen. Each bond is a covalent bond due to chain transfer of an olefin monomer double bond by radical generation by proton abstraction of a hydroxyl group of a water-soluble linear polysaccharide cationic derivative. In this reaction, the product occurs in latex. This latex polymer is generally insoluble in water, alcohol, or an organic solvent such as acetone or tetrahydrofuran, but by a casting method or the like,
Easy film formation. Alternatively, an excessively insoluble solvent such as alcohol is added to obtain a precipitate, and then a molded product can be easily formed by a hot press method or the like. For the above purpose, the ratio of the trunk polymer to the graft polymer or the polymerization degree ratio thereof in the graft polymer can be variously selected according to the purpose. In the graft polymerization, the polymerization rate is determined by the graft rate (%). This is the graft ratio (%) = (amount of monomer graft-polymerized / amount of trunk polymer in graft copolymer)
It is defined as × 100. In the present invention, the olefin compound is formed as a graft chain, and a graft ratio in the range of 2% to 5000% is considered appropriate. It has been repeatedly stated that the present invention is a product obtained by graft-polymerizing an olefin monomer underwater into a cationic derivative of a water-soluble linear polysaccharide having a hydroxyl group, but the structure of the resulting copolymer chain is claimed. As described as the chemical structural formula in the range of, the formula is composed of ( Chemical Formula 2) and ( Chemical Formula 3 ) and is represented by the following ( Chemical Formula 1).

## STR00001 ## Each bond is a covalent bond due to chain transfer of an olefin monomer double bond due to radical generation by abstraction of a proton of a hydroxyl group of a cationic derivative of a water-soluble linear polysaccharide.

[0006]

Example 1 2 g of DEAE (diethylaminoethyl) dextran hydrochloride having a nitrogen content of 5% with dextran having an average molecular weight Mw of 500,000 as a matrix was dissolved in 50 ml of water, and then 5 ml of methanol and 30 ml of methyl methacrylate (MMA) were added. In addition, the reaction solution and air in the reaction vessel were sufficiently replaced with nitrogen gas, and then the mixture was thoroughly stirred, and the solution was dissolved in 15 ml of 0.1N nitric acid.
Add mg to start the reaction. The reaction is carried out at 30 ° C. for 1 hour to form a latex. At the end of the reaction, 3 ml of a 1% solution of hydroquinone was used as a terminator. Then, the reaction solution was poured into 3 times the amount of methanol to obtain a precipitate. The precipitate was thoroughly washed with hot water, centrifuged and dried under reduced pressure at 50 ° C., and the dried product was placed in a Soxhlet extractor and subjected to acetone extraction for 24 hours to obtain DEAE (diethylaminoethyl) dextran-MMA copolymer hydrochloric acid. 1.5 g of salt was obtained. Nitrogen content 2.0% Graft ratio 25% vs. DEAE-dextran yield 30% This is insoluble in water, which is a good solvent for DEAE dextran hydrochloride, and acetone, which is a good solvent for polymethylmethacrylate. In the infrared absorption spectrum of this product, absorption of a carbonyl group, which is not found in DEAE-dextran hydrochloride, is found near 1730 cm ⁻¹ . Example 2 After carrying out the same reaction as in Example 1, the reaction-completed solution of the latex was not poured into methanol, but the solution was allowed to undergo transposition in water to remove unreacted materials and an initiator, and a homopolymer was mixed. A DEAE dextran-MMA copolymer latex was obtained. This is useful as a reagent for detecting rheumatoid factor and shows the test results. Test Method An equal amount of antibody in a 2% solution of this latex [ie 2
Glycine-Nac so as to become 0 μg / mg latex
l buffer-γ globulin (GB) (0.05 mol /
1, diluted with PH 8.2)] while stirring 1
Add dropwise and stir at room temperature for 2 hours, then (400
Centrifuge at 0 rpm for 30 minutes and repeat resuspension with GB)
Finally at GB with 0.5% BSA (bovine albumin) 1
Store at 4 ° C as a% suspension. A drop of the sensitized latex liquid of the present invention was dropped on a wiped glass plate with a capillary tube,
Rheumatoid factor-positive serum diluted 1/20 with GB in advance was added dropwise to a 0.05 ml latex solution on a glass plate with a capillary tube, stirred and shaken. When judged, aggregates were found. When the same operation was performed with a known 1/20 diluted rheumatoid factor negative serum, no aggregate was observed. Example 3 4 g of DEAE (diethylaminoethyl) -pullulan hydrochloride having a nitrogen content of 4% and having pullulan having an average molecular weight Mw of 200,000 as a matrix was dissolved in 80 ml of water, and then 10 ml of methanol was added.
ml, 35 ml of styrene monomer were added, the reaction solution and the air in the reaction vessel were sufficiently replaced with nitrogen gas, and then 200 mg of ceric ammonium nitrate nitrate dissolved in 30 ml of 0.1N nitric acid was added with thorough stirring. Start the reaction. The reaction is carried out at room temperature for 1 hour to form a latex. At the end of the reaction, 3 ml of a 1% solution of hydroquinone was used as a terminator. The subsequent purification and drying steps were performed in the same manner as in Example 1 to obtain 6 g of DEAE (diethylaminoethyl) pullulan-styrene copolymer hydrochloride. Nitrogen content 1.14% Grafting ratio 350% vs DEAE pullulan yield 43% Example 4 After the same reaction as in Example 3, the reaction completed solution of the latex was not poured into methanol, and the solution was subjected to folding in water. After that, the unreacted material and the initiator were removed to obtain a DEAE pullulan-styrene copolymer latex in which a homopolymer was mixed. This was useful as a reagent for detecting rheumatoid factor. As in Example 2, a 2% solution of this latex in an equal amount of the antibody, [ie glycine-Nacl buffer-γ globulin (GB) (0.05 mol / l, PH 8.2) to give a 20 μg / mg latex).
The latex solution diluted with and sensitized with a rheumatoid factor-positive serum was visually judged by the procedure of Example 2 for the presence or absence of agglutination, but agglutinates were observed in the rheumatoid factor-negative serum. There wasn't. Example 5 4 g of AE (aminoethyl) dextran hydrochloride having a nitrogen content of 5% and having dextran having an average molecular weight Mw of 40,000 as a matrix
Was dissolved in 90 ml of water, then 5 ml of methanol and 20 ml of butyl methacrylate were added, and the air in the reaction solution and the reaction vessel was sufficiently replaced with nitrogen gas. 50 mg of cerium ammonium nitrate is added to start the reaction. The reaction is carried out for 30 minutes at room temperature to form a latex.
When the reaction ends, 3m of hydroquinone 1% solution is used as a terminating agent
1 was used. Subsequent purification and drying steps were performed in the same manner as in Example 1 to obtain 6 g of hydrochloride of AE (aminoethyl) dextran-butyl methacrylate copolymer. Nitrogen content 2.0% Graft ratio 300% AE-dextran yield 30% This is insoluble in water, which is a good solvent for AE dextran hydrochloride, and acetone, which is a good solvent for polybutyl methacrylate. Example 6 After performing the same reaction as in Example 5, the reaction-completed solution of the latex was not poured into methanol, but the solution was allowed to disintegrate in water to remove the unreacted materials and the initiator, and the homopolymer was mixed. An AE (aminoethyl) dextran-butyl methacrylate copolymer latex was obtained. This was useful as a reagent for detecting rheumatoid factor. As in Example 2, a 2% solution of this latex in an equal amount of the antibody [ie glycine-to give 20 μg / mg latex-
Nacl buffer-γ globulin (GB) (0.05 m
The latex solution diluted with ol / l, PH 8.2) and sensitized was visually evaluated for the presence or absence of aggregation in the rheumatoid factor positive serum according to the procedure of Example 2. No aggregates were found in the negative serum. Example 7 Nitrogen content 3 with pullulan having an average molecular weight Mw of 30,000 as a matrix
% HPTMA (2-hydroxypropyltrimethylammonium) -pullulan hydrochloride 4 g was dissolved in 100 ml water, then 30 ml methyl acrylate monomer was added,
After sufficiently replacing the air in the reaction solution and the reaction vessel with nitrogen gas, while stirring well, cerium ammonium nitrate nitrate 20Omg ceric nitrate dissolved in 20 ml of 0.1N nitric acid
To start the reaction. The reaction is carried out at room temperature for 1 hour to form a latex. At the end of the reaction, 4 ml of a 1% hydroquinone solution was used as a terminator. Subsequent purification and drying steps were performed in the same manner as in Example 1 to obtain 2 g of hydrochloride of HPTMA (2-hydroxypropyltrimethylammonium) pullulan-methyl acrylate copolymer. Nitrogen content 2% Graft ratio 150% HPTMA pullulan yield 33% Example 8 After the same reaction as in Example 7, the reaction was carried out in water to remove unreacted materials and initiator to obtain a homopolymer. A mixed HPTMA (2-hydroxypropyltrimethylammonium) pullulan-methyl acrylate copolymer latex was obtained. This was useful as a reagent for detecting rheumatoid factor. As in Example 2, a 2% solution of this latex in an equal amount of antibody [ie 20 μg / m 2
Glycine-Nacl buffer-γ globulin (GB) (0.05 mol / l, PH
The latex liquid diluted with 8.2) and sensitized was prepared in Example 2
When the presence or absence of agglutination was visually determined in the rheumatoid factor-positive serum by the above procedure, an agglutinate was observed, but no agglutinate was observed in the rheumatoid factor-negative serum.

Claims (2)

[Claims] 1. The formula of the unit of the cationic derivative of the water-soluble linear polysaccharide is [C6 H7 O2 (OH) 3-a. (OX) a] x.H2 O (1) [wherein X is -(CH2) nR1 (R1 is -NH
3 +, -NH + (CH3) 2, -NH +
(C2 H5) 2, -N + (CH2) 2CH2
CH (OH) CH3, -N + (C2H5) 2
(C2H5) N (C2H5) 2, -N +
(C2 H5) 2 CH2 CH (OH) CH3,
-N + (C2H5) 3, -C6H4.N
H3 +, -CO · C6 H4 · NH3 +, a group selected from the group consisting of n = an integer of 1 to 3) or -CO
R2 (R2 is -CH2.NH3 + or -C6
H4 · NH3 +) or —CH2 CH (OH) ·
CH2 R3 (R3 is -NH3 +, -NH +
(CH3) 2, -NH + (C2H5) 2,
-N + (a group selected from the group consisting of C2H5) 3) or -NH.CH2.CH2, a is 0 <a <3.
Is a positive number, x is an integer of 50,000 ≧ x ≧ 5 [Wherein R4, R5 and R6 are each a hydrogen atom or CH3 C12 alkyl group, cyclohexyl group, C1 to C4
Lower alkyl-substituted cyclohexyl group, C1-C8
Hydroxyalkyl group, C1-C8 aminoalkyl group, C1-C8 dialkylaminoalkyl group, glycidyl group, tetrahydrofuran group, C1-C
4, a lower alkyl-substituted tetrahydrofuran group, a benzyl group and (-CH2CH2-O-) yCH2CH.
2 OH group (where y is a positive number of 1 to 10), -N (R9
) 2 (R9 is a hydrogen atom or a C1-C4 alkyl group, two R9 may be the same or different), (R10 is a C1 to C8 alkyl group), phenyl group, pyridine group, tolyl group, pyrrolidone group and C1 to
C4 represents a lower alkyl-substituted pyrrolidone group]. m is 2
It is a polymer of an olefin compound represented by a positive integer of 0 to 200,000. A cation derivative of water-soluble linear polysaccharide is used as a stem polymer, and its hydroxyl group is used as a binding site by abstracting a hydrogen atom, and an olefin compound is used as a graft chain. The graft ratio is 2% to 5000%.
The following (3) consisting of the equation (1) and the equation (2) in the range of
[[C6H7O2 (OH) 3-a. (OX) a] x.H2O-]-[-R4CR5R6CR7-] m (3) Water-soluble linear polysaccharide is used as a matrix. Water-soluble Linear Polysaccharides Obtained by Grafting Olefin Monomers on Cationic Partial Substitutes of Polysaccharides Copolymers Obtained by Grafting Olefin Monomers on Water in Cationic Partial Substitutes of Polysaccharides latex. 2. The formula of the unit of the cationic derivative of the water-soluble linear polysaccharide is [C6 H7 O2 (OH) 3-a. (OX) a] x.H2 O (1) [wherein X is -(CH2) nR1 (R1 is -NH
3 +, -NH + (CH3) 2, -NH +
(C2 H5) 2, -N + (CH2) 2CH2
CH (OH) CH3, -N + (C2H5) 2
(C2H5) N (C2H5) 2, -N +
(C2 H5) 2 CH2 CH (OH) CH3,
-N + (C2H5) 3, -C6H4.N
H3 +, -CO · C6 H4 · NH3 +, a group selected from the group consisting of n = an integer of 1 to 3) or -CO
R2 (R2 is -CH2.NH3 + or -C6
H4 · NH3 +) or —CH2 CH (OH) ·
CH2 R3 (R3 is -NH3 +, -NH +
(CH3) 2, -NH + (C2H5) 2,
-N + (a group selected from the group consisting of C2H5) 3) or -NH.CH2.CH2 |, a is 0 <a <3.
And x is an integer of 50,000 ≧ x ≧ 5]. [Wherein R4, R5 and R6 are each a hydrogen atom or CH3 C12 alkyl group, cyclohexyl group, C1 to C4
Lower alkyl-substituted cyclohexyl group, C1-C8
Hydroxyalkyl group, C1-C8 aminoalkyl group, C1-C8 dialkylaminoalkyl group, glycidyl group, tetrahydrofuran group, C1-C
4, a lower alkyl-substituted tetrahydrofuran group, a benzyl group and (-CH2CH2-O-) yCH2CH.
2 OH group (where y is a positive number of 1 to 10), -N (R9
) 2 (R9 is a hydrogen atom or a C1-C4 alkyl group, two R9 may be the same or different), (R10 is a C1 to C8 alkyl group), phenyl group, pyridine group, tolyl group, pyrrolidone group and C1 to
C4 represents a lower alkyl-substituted pyrrolidone group]. m is 2
It is a polymer of an olefin compound represented by a positive integer of 0 to 200,000. A cation derivative of a water-soluble linear polysaccharide is used as a stem polymer, and its hydroxyl group is used as a binding site by abstracting a hydrogen atom, and an olefin compound is used as a graft chain. The graft ratio is from 2% to 5000.
[[C6H7O2 (OH) 3-a. (OX) a] x.H2O-]-represented by the following formula (3) consisting of the formula (1) and the formula (2) in the range of%. [-R4 CR5 R6 CR7-] m (3) Cationic property of water-soluble linear polysaccharide obtained by grafting an olefin monomer onto a cationic partial substitution product of a polysaccharide having a water-soluble linear polysaccharide as a base A latex diagnostic agent obtained by adsorbing an antibody to a latex made of a copolymer obtained by grafting an olefin monomer on a partially substituted product in water.