JPS62153751A - Absorber carrier for chromatography - Google Patents

Abstract

PURPOSE:To obtain a desirable affinity gel, by constituting said gel of a copolymer based on glycidyl monovinyl ester A and alkylene glycol divinyl ester B and containing protein bonded to the epoxy group of the A-component. CONSTITUTION:In a gel-like copolymer wherein (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) alkylene glycol divinyl ester and main components and the (A)-component is crosslinked with the (B)-component, protein A is bonded to the epoxy group of the (A)-component through a covalent bond to prepare a porous copolymer used as the titled adsorbing carrier. This gel-like copolymer can be prepared by obtaining a porous gel through the aqueous suspension polymerization of the (A)-component and the (B)-component in the presence of a water-insoluble org. diluent.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an adsorption carrier for chromatography that can be used in various chromatography including affinity chromatography.

Affinity chromatography is a type of chromatography technology that uses the affinity of a pair of substances to specifically interact with each other to separate and purify biological substances. It is useful for identifying and purifying chemical properties, that is, a specific chemical structure on a molecule.

An adsorption carrier (affinity gel) for affinity chromatography is an active support obtained by bonding a binding group (spacer) to an insoluble carrier (matrix), and a ligand is bonded to the spacer. An adsorption operation is performed by selecting a combination of substances that interact with each other.

As one of the ligands, protein A is 5taphyl
This protein is obtained from the cell wall of Ococcus aureus and has the property of strongly adsorbing to immunoglobulin, so affinity chromatography using protein A as a ligand is considered to be useful as a means of research related to the immune system. It is being

For example, in separation, purification, and analysis by affinity chromatography in which protein A is used as a ligand and IgG is used as the adsorption target substance, the desired properties of the active support, which is the main component of the adsorption carrier for affinity chromatography, are as follows: Low non-specific adsorption;
It has high porosity, easy binding of lycand and large immobilization capacity, chemical stability and pH range,
It is sufficiently stable and does not change in volume under a wide range of conditions including 4 degrees of salt and temperature, has sufficient mechanical strength and stability, has good flow properties, and is resistant to biological contamination. .

Cellulose, dextran, polyacrylamide, agarose, and the like, which have been conventionally used as base materials for adsorption carriers for affinity chromatography, do not necessarily have these desired properties. In particular, since it is a so-called soft gel lacking in hardness, it has serious drawbacks such as poor flow properties and poor separation properties, and also has a short lifespan.

Furthermore, although the silica beads that have been used in recent years are satisfactory in terms of hardness, they cannot be used under alkaline conditions, which poses the problem of placing significant restrictions on the selection of separation conditions and elution/washing conditions. Ta.

SUMMARY OF THE INVENTION An object of the present invention is to overcome the drawbacks of the conventional adsorption carriers for chromatography, and fully possess the above-mentioned properties desired in an adsorption carrier for affinity chromatography. An object of the present invention is to provide an adsorption carrier for chromatography.

Means for Solving the Problems The present invention provides an adsorption carrier for chromatography that can achieve the above objects.

That is, the present invention comprises (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) alkylene glycol divinyl ester as main components;
The component is a gel-like copolymer crosslinked with component (B) (hereinafter referred to as
A porous copolymer containing protein A covalently bonded to an epoxy group via a bonding group based on the component (A) of the gel-like copolymer according to the present invention. The present invention relates to an adsorption carrier for chromatography.

The adsorption carrier for chromatography of the present invention will be explained below.

The gel-like copolymer according to the present invention is disclosed in JP-A-60-1042
As described in Publication No. 56, for example, (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) alkylene glycol divinyl ester are mixed in an aqueous solution in the presence of a water-insoluble organic diluent. It can be prepared by turbid polymerization to obtain a porous gel.

Since the components (A) and (B) are sparingly soluble in water, they can be copolymerized by an aqueous suspension polymerization method, and the aqueous suspension polymerization can be carried out by a simple N oil-in-water suspension method. . Copolymerization is carried out in the presence of an organic diluent, and due to the presence of this organic diluent, the alkylene glycol divinyl ester component (B) acts as the main crosslinking component, and the glycidyl monovinyl ester component (A) or Most of the epoxy groups in the glycidyl monovinyl ether component remain in the resulting copolymer without ring opening, and are also useful for adjusting the pore size of the resulting porous gel.

The component (A) is glycidyl ester of monovinylcarboxylic acid having 3 to 12 carbon atoms or 3 to 12 carbon atoms.
Glycidyl ether of monovinyl alcohol is used. Among these, those having a small number of carbon atoms are particularly preferably used, and examples thereof include glycidyl methacrylate, glycidyl acrylate, allyl glycidyl ether, and the like.

On the other hand, the (B) component that acts as a crosslinking component has 2 carbon atoms.
Or esters of alkylene glycols or polyalkylene glycols of 3 and acrylic acid or methacrylic acid are preferably used.

Examples include alkylene glycol divinyl esters such as ethylene glycol diacrylate, ethylene glycol dimethacrylate, propylene glycol diacrylate, propylene glycol dimethacrylate, and polyalkylene glycol divinyl esters such as polyethylene glycol dimethacrylate, polybrobylene glycol dimethylate. .

The ratio of component (A) to component (B) is 10 to 90 mol% for the former and 90 to 10 mol% for the latter.

Preferably (A) component 40 to 80 mol%, (B) component 6
It is 0 to 20 mol%. (A) Less than half of the component is a comonomer, namely methyl methacrylate, methyl acrylate,
Lower vinyl esters such as vinyl acetate can be substituted. Of course, the greater the proportion of component (A), the greater the content of epoxy groups in the copolymer. The higher the proportion of component (B), the higher the degree of crosslinking of the copolymer, and therefore the denser the network structure, the lower the degree of swelling, and the harder the copolymer becomes. If the ratio of both components is outside the above range, a gel-like copolymer that fully satisfies the desired characteristics and properties cannot be obtained.

The organic diluent to be used may be one that is inert to the polymerization reaction, insoluble or sparingly soluble in water, but capable of dissolving the additives in the raw material. The amount used is at least 30% by volume based on the total amount of monomer components and organic diluent, preferably 40-8% by volume.
0% by volume. The larger the amount used, the more suppressed is the ring opening of the epoxy group during the polymerization process. Generally speaking, use (A
) It is possible to easily make about 60 to 95% of the epoxy groups contained in the charged amount of the component remain in the produced polymer.

It is believed that the remainder is hydrolyzed during the polymerization process and a portion is used for crosslinking.

As mentioned above, the content of epoxy groups in the resulting gel can be adjusted by the amount of component (A) used, i.e. the ratio to component (B), or by partial substitution with a comonomer, and also by the amount of organic diluent. Preferably, the epoxy oxygen is present in the resulting polymer in an amount of 1% by weight or more, expressed as epoxy oxygen, up to about 10% by weight.

As mentioned above, the use of organic diluents during aqueous suspension polymerization also helps control the pore size of the resulting porous gel. When an organic diluent having a low swelling property is used for the resulting gel, phase separation occurs in the suspended particles during the polymerization process, and as a result, a gel with a large pore size, that is, a gel having a so-called giant pore structure can be obtained. On the other hand, if a highly swelling diluent is used, the polymerization will proceed in a swollen state during the polymerization process.
Phase separation is difficult to occur and a polymer with relatively small pores formed by swelling is obtained. The degree of swelling can be determined based on the solubility parameter of the organic diluent. Examples of organic diluents that can be suitably used in the present invention include cyclohexanone, chlorohenzene, henzene, toluene, and
n-propyl acetate, n-butyl acetate, n-
Octane etc.

The aqueous suspension polymerization can be carried out in the presence of a free-radical generating catalyst according to conventional methods known per se.

The amount of the aqueous phase is approximately the same volume or more based on the amount of the organic phase, and although there is no particular restriction, it may be used in an amount up to about 10 times the volume.

The introduction of the bonding group into the gel-like copolymer according to the present invention includes:
This can be carried out by utilizing the reactivity of the epoxy group exposed to the component (A) of the gel-like copolymer according to the present invention, but in this case, the binding group has a functional group capable of covalently bonding to protein A. However, there is no particular restriction on the structure of the bonding group other than having the above-mentioned functional group. Examples of the functional group capable of covalently bonding with protein A include an epoxy group, an amino group, a hydrazino group, a carboxyl group, a formyl group, and a thiol group.

In this case, the bonding group can be easily introduced using a known method, and it does not necessarily have to be introduced in one reaction step, but can also be introduced in two or more reaction steps. . A specific example of introducing a bonding group will be described below.

(i) Introduction of a bonding group having an epoxy group as a functional group For example, the epoxy group based on the component (A) of the gel copolymer according to the present invention is generated by ring-opening modification with a hydroxyl group-containing compound such as water. A compound having a glycidyl group is bonded to a hydroxyl group to introduce a bonding group having an epoxy group based on a glycidyl group. (Hereinafter, a polymer into which a glycidyl group has been introduced in this way will be referred to as a modified gel copolymer according to the present invention.) Ring-opening modification is performed in the presence of an acid or an alkali,
For example, when carrying out in the presence of an acid, these copolymers are heated at 70°C to 1°C in water using sulfuric acid or hydrochloric acid as a catalyst.
This is done by heating to 00°C for about 2 hours. The concentration of the acid is preferably 0.1N to 0.5N. Ring-opening modification is performed almost quantitatively.

Compounds having a glycidyl group that are bonded to the hydroxyl group generated by ring-opening modification include epihalohydrins such as shrimp chlorohydrin and shrimp chlorohydrin/, ethylene glycol diglycidyl ether, and 1,4-butanediol. Diglycidyl ether, polyethylene glycol jig! Examples include diglycidyl ethers such as diyl ether, and alkyl diene diepoxides such as 3-butadiene diepoxide and 1,7-octazidine diepoxide.

In order to act with a compound having a glycidyl group, for example, a ring-opening modified gel copolymer is reacted with shrimp chlorohydrin in an aqueous solution of an inorganic base such as 1-lium hydroxide or potassium carbonate, or A hydrophilic copolymer is reacted with ethylene glycol or 1,4-butanediol diglycidyl ether, or polyethylene glycol diglycidyl ether in an aqueous solution of an inorganic base such as sodium hydroxide or potassium carbonate containing sodium borohydride. etc. Repeating unit of polyethylene glycol diglycidyl ether used -C2
8a It is preferable to use one in which the number of O- repeats is 1 to 10.

(ii) Introduction of a bonding group having an amino group, hydrazino group, carboxyl group, etc. as a functional group For example, as a method for introducing a bonding group having an amino group, hydrazino group, carboxyl group, etc., the gel copolymer according to the present invention may be used. Based on the epoxy group based on component (A) or the glycidyl group of the modified gel copolymer according to the present invention (epoxy group), the general formula (I) H!NR' (wherein R' is a hydrogen atom, (representing an amino group, an ω-aminoalkyl group, or an ω-carboxyalkyl group. is further treated with an alkylenecarboxylic anhydride to form the epoxy group or glycidyl group with the general formula (II) NHR (wherein R is a hydrogen atom, an amino group, an ω-aminoalkyl group, an ω-carboxyalkyl group, ω-carboxyalkanoyl group, ω-carboxyalkanoylamino group or ω
- represents a carboxyalkanoylaminoalkyl group.

) There is a method of introducing a bonding group represented by

In this case, the reaction can be carried out according to the following reaction formula.

In addition, in the following reaction formula, (base 10CH-CH2 is ゛・,
1 represents a gel copolymer according to the present invention or a modified gel copolymer according to the present invention, and -CH-CH2 represents one epoxy group or glycidyl group in these copolymers. In the formula, m is 1 or 2, and n, p, q and r are each an integer of 1 or more (more preferably 1 to 12).

-m-self/ On the surface, the reaction formula (D), -1 formula (D ') or the compound of general formula (F) can be carried out in the presence of a solvent, but if these compounds are liquid under the reaction conditions, no solvent may be used. As a solvent, water is usually used, but other hydrocarbons such as n-hexane, benzene, and quinolene, tonoethers such as tetrahydrofuran and dioxane, chlorinated hydrocarbons such as chloroform and chlorohenzene, and alcohols such as ethanol and methyl cellosolve are also used. In addition to ketones such as acetone and methyl isobutyl ketone, dimethylformamide and dimethyl sulfoxide are also used, and these solvents may be used alone or in a mixed solvent system. Although no particular catalyst may be used, hydroxides or carbonates of alkali or alkaline earth metals such as sodium hydroxide and potassium carbonate may also be used.

Examples of the compound of general formula (D') include ethylenediamine,
1,3-diaminopropane, 1,4-diaminobutane,
1,5-diaminopentane, 1,6-diamithexane, 1,7-diaminohbutane, 1,8-diaminooctane, 1,9-diaminononane, 1,10-diaminodecane, 1,12-diaminododecane, etc. diaminoalkanes.

Examples of the compound of general formula (F) include aminocarboxylic acids such as glycine, β-alanine, 4-aminobutyric acid, 6-aminocaproic acid, and 8-aminocaprylic acid.

The reaction conditions are not necessarily limited, but it is generally preferable to select the following conditions to carry out the reaction.

Weight (a) and formula (D) and general formula (D') of the gel-like copolymer according to the present invention or the modified gel-like copolymer according to the present invention
Or the ratio of the weight (b) of the compound of general formula (F): a: b= 1: 0.3-10, more preferably a: b= l: 0.3-3, reaction temperature 20-150°C, More preferably room temperature to 100
C, reaction time: 1 to 60 hours, more preferably 1 to 30 hours, reaction pressure normal pressure to 10 atm, more preferably normal pressure.

There are no special requirements for post-treatment after the reaction, and conventional methods such as separation and washing may be carried out as appropriate.

The chromatography carrier according to the present invention obtained above can be prepared using an active support group as already shown in the general formula, if necessary, when the compound of formula (D) or general formula (D') is used. The terminal amino group of can be converted into an active supporting group having a carboxyl group by reacting with the acid anhydride of general formula (E). Water is usually used as a solvent for the reaction, but ethers such as tetrahydrofuran and dioxane, carboxylic acids such as acetic acid, pyridine, and the like may also be used. Further, although a catalyst may not be used, an acid such as hydrochloric acid or sulfuric acid or an alkali such as sodium hydroxide or potassium carbonate is added for 30 hours, and the reaction pressure is normal pressure to 10 atm, more preferably normal pressure.

There are no special requirements for post-treatment after the reaction, and it may be appropriately carried out by commonly used methods such as separate furnaces and washing.

In addition, as another method for introducing a bonding group having a carboxyl group as a functional group, an aminocarboxylic acid having a hydroxyl group is added to the component (A) of the gel copolymer according to the present invention (
There is a method of binding by acting on the epoxy group or the glycidyl group of the modified gel copolymer according to the present invention.

Examples of aminocarboxylic acids having a hydroxyl group include serine, homoserine, threonine, 4-hydroxyproline, 4-
Examples include amino acid derivatives such as amino-3-hydroxybutyric acid, N-tris(hydroxymethyl), and methylglycine, and amino sugar derivatives such as glucosaminic acid.

The bonding state is as a result of the bonding between the epoxy group of the gel copolymer according to the present invention or the glycidyl group of the modified gel copolymer according to the present invention with the amine group of the aminocarboxylic acid having a hydroxyl group. The glycidyl group opens to form a hydroxyl group and a carboxyl residue having a hydroxyl group.

The action of aminocarboxylic acid with 11M of water is as follows:
It can be carried out in the presence of a solvent.

Water is usually used as a solvent, but other solvents include hydrocarbons such as n-hexane, benzene, and xylene, ethers such as tetrahydrofuran and dioxane, chlorinated hydrocarbons such as chloroform and chlorohenzene, and ethanol and methyl cellosolve. In addition to alcohols and ketones such as acetone and methyl isobutyl ketone, dimethylformamide and dimethyl sulfoxide are also used, and these solvents may be used alone or in a mixed solvent system.

Although no particular catalyst may be used, hydroxides or carbonates of alkali or alkaline earth metals such as sodium hydroxide and potassium carbonate may also be used.

There are no particular limitations on the reaction conditions (for example, the same conditions as in the case where the compound of general formula (1) is allowed to act) can be used.

When a bonding group is introduced using an aminocarboxylic acid having such a hydroxyl group, nonspecific adsorption originating from hydrophobicity based on a portion other than the base material, that is, a portion of the bonding group can also be eliminated.

(iii) Introduction of a bonding group having a formyl group as a functional group For example, the bonding group having an epoxy group as a functional group obtained in the above (i) can be easily hydrolyzed into 1
, 2-glycol, and this 1.2-
Glycol can be converted into formyl group by the action of periodic acid. There are no particular restrictions on the reaction conditions, and the reaction can be carried out according to a conventional method using, for example, water, ethanol, acetic acid, or a mixture thereof as a solvent at room temperature to 50° C. by reacting periodic acid or an inorganic salt thereof. .

(iv) Introduction of a bonding group having a thiol group as a functional group For example, the bonding group having an epoxy group as a functional group obtained in (1) above is prepared by treating the functional group with sodium thiosulfate and then adding hydrochloric acid. A thiol group can be introduced by, for example, reacting with a thiol group.

There are no particular restrictions on the reaction conditions, and the reaction can be easily carried out according to a conventional method, for example, at room temperature in an aqueous solvent.

When protein A is covalently bonded to a binding group, it can be carried out in an appropriate solvent using an appropriate catalyst or reaction reagent, if necessary depending on the functional group possessed by the binding group. For example, as catalysts, acids such as hydrochloric acid, sodium carbonate, or sodium bicarbonate, and alkalis are mainly used when the functional group is an epoxy group, and as reaction reagents, for example, N-hydroxysuccinimide and dicyclohexylcarbodiimide are used. When the functional group is a carboxyl group,
Also, 1-ethyl-3-(3-dimethylaminopropyl)
A condensing agent such as carbodiimide is used when the functional group is a carboxyl group, an amino group or a hydrazino group, a reducing agent such as sodium cyanoborohydride is used when the functional group is a formyl group, etc. I can do it. Water is usually used as a solvent, but if necessary, a warm solution of phosphoric acid or acetic acid may be used, or an inorganic salt such as sodium chloride may be added.

The conditions for the reaction with protein A are not necessarily limited, but it is generally appropriate to carry out the reaction within the following range.

Weight ratio of the gel-like copolymer or modified gel-like copolymer according to the present invention to which a binding group is attached and protein A: 1:0.0
3 to 0.3, preferably 1:0.05 to 0.2; Reaction temperature: 0°C to room temperature, preferably 4°C to room temperature; Reaction time: 1
~72 hours, preferably 2-12 hours.

There are no special requirements for post-treatment after the reaction;
Washing and the like are carried out as appropriate using commonly used methods.

Effects of the Invention The adsorption carrier for chromatography provided by the present invention uses a base having all the properties desired as an adsorption carrier for affinity chromatography as described above, and binds protein A to this base through an appropriate bonding group. Since they are commonly bonded, by using the adsorption carrier of the present invention,
Not only can it be used as a packing material for affinity chromatography in the same way as adsorption carriers using conventional soft gels (in particular, it can be packed into pressure-resistant columns and used under pressure, which means that the working time can be reduced). Since the width can be shortened to a large extent, the greatest advantage is that the efficiency of separation and purification can be greatly improved. It goes without saying that it is essential for measurement.

In particular, while the gel copolymer has many advantages as described above, adsorption carriers to which Protein A is covalently bonded via the bonding group as described above, such as those used for -S, are Compared to adsorption carriers for affinity chromatography using cyanogen oxide, the adhesive does not fall off during use, and the length of the bonding group can be adjusted arbitrarily, making it possible to adsorb the ligand to the target substance. The adsorption carrier for chromatography provided by the present invention is highly effective in that it has excellent performance and low specific adsorption.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Representative examples of the method for producing the adsorption carrier for chromatography of the present invention from known substances will be shown below and explained in more detail. However, these are merely examples for explanation, and it goes without saying that the present invention is not limited thereto.

Example 1 An epoxy group-containing gel copolymer obtained from glycidyl methacrylate and ethylene glycol dimethacrylate was subjected to ring-opening modification of the epoxy group with water, and further epoxy groups were introduced with 1,4-butanediol diglycidyl ether. 1.0 g of gel (epoxy groups: 0.3 mmol/g dry gel) was added to a solution of Protein A20 in 0.05 molar sodium bicarbonate and 0.05 molar sodium carbonate aqueous solution in 1 ml and shaken for 8 hours at room temperature. rear,
It was left at 4°C overnight. After the gel was renewed and washed with a 1M aqueous sodium chloride solution and water, 1M ethanolamine 21I11 was added and shaken at room temperature for 1 hour, and the gel was renewed again and washed with water. It was confirmed from the amount of unreacted recovered protein that the adsorption carrier thus obtained supported 2.2 rrg of protein A per 1 g of dry gel.

Example 2 1.0 g of gel into which epoxy groups were introduced in Example 1
Sodium periodate 1. Acetic acid water containing Og (9:
1) Added to 20ml of solution and shaken at room temperature for 2 hours. After the gel was renewed and washed with water, it was added to 3 ml of aqueous i'J solution containing 0.03 mol acetic acid and 0.06 mol sodium acetate, and 20 ml of protein A was added thereto, followed by shaking at room temperature for 8 hours. After the gel was separated and washed with a 1 molar aqueous sodium chloride solution and water, it was added to 20 μl of sodium cyanoborohydride in 2 ml of 1 molar Tris salt solution (pH 8,0)/8 solution at room temperature. After shaking for 1 hour, the gel was harvested again and washed with water. It was confirmed that the adsorption carrier thus obtained supported 9.7 μg of protein A per 1 g of dry gel.

Example 3 An epoxy group-containing gel copolymer obtained from glycidyl methacrylate and ethylene glycol dimethacrylate was subjected to ring-opening modification of the epoxy group with water, and then, after introducing an epoxy group with ethylene glycol diglycidyl ether, further After thoroughly washing 1.0 g of gel into which carboxyl groups were introduced with 4-aminobutyric acid with anhydrous dioxane, it was added to 4 ml of anhydrous dioxane, and further 80 μ of N-hydroxysuccinimide and 144 μ of dicyclohexylcarbodiimide were added. After shaking for 2 hours at room temperature, the gel was separated and added with 20 m of anhydrous dioxane. ! , methanol 6 ml
and 3 ml of cold water.

This gel was added to a solution of 0.01 mol of Protein A20 and sodium bicarbonate in 2 ml of water, shaken at room temperature for 2 hours, and then left overnight at 4°C. After the gel was renewed and washed with a 1M sodium chloride aqueous solution and water, it was added to 2 mj2 of 1M Tris-HCl buffer (pH 8,0) and shaken at room temperature for 1 hour, and then the gel was renewed again and washed with water.

It was confirmed that the adsorption carrier thus obtained supported 12.5 mg of protein A per 1 g of dry gel.

Test Example 1 The adsorption carrier obtained in Example 2 had a diameter of 4.6 mm and a length of 751 mm.
When Hill-1gG was analyzed using a high-performance liquid chromatography device, the first
A romatogram was obtained as shown in the figure. Analysis conditions: Eluent 020.1 molar sodium phosphate buffer (pH 4,5
), eluent 020.1 molar sodium phosphate buffer (p
H3,0), elution rate 1.0 mj! /min detector ultraviolet spectrophotometer 280 nm.

Figure 1 shows that the human IgG used here does not adsorb under the pH 3 condition (Figure 1 (A)), but it adsorbs and elutes 2, and it completely adsorbs under the pH 4 and 5 conditions. This shows that (Figure 1 (B)).

Test Example 2 In Test Example 1, the same procedure was carried out using the adsorption carrier obtained in Example 3 instead of the adsorption carrier obtained in Example 2, and using bovine T-globulin instead of human gG. When the analysis was performed, a chromatogram as shown in Figure 2 was obtained.

Figure 2 shows that the bovine γ-globulin provided here is not adsorbed under the condition of pH 3 (Figure 2 (A)) or the adsorbed fraction is also eluted, whereas under the condition of pH 4 and 5, the adsorbed fraction and the adsorbed fraction are eluted. This shows that a fraction that is not adsorbed is included (Figure 2 (B)).

[Brief explanation of drawings]

FIG. 1 and FIG. 2 are both chromatograms showing patterns when adsorption and elution of globulin were investigated using the adsorption carrier for chromatography according to the present invention. In both Figures 1 and 2, (A) shows the case where only eluent ■ is used, and (B) shows the case where eluent ■ was initially used and then switched to eluent ■ at the arrow. .

Claims (1)

[Claims] Epoxy based on component (A) of a gel-like copolymer containing (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) alkylene glycol divinyl ester as main components, and component (A) crosslinked with component (B). An adsorption carrier for chromatography comprising a porous copolymer containing protein A covalently bonded to a base via a bonding group.