JPS62161052A - Adsorbing carrier for chromatography - Google Patents

Abstract

PURPOSE:To improve the separation characteristic of chromatography, by using a copolymer, which is obtained by bonding phenyl borate to a specific gel like copolymer based on glycidyl monovinyl ester or the like through a bonding group, as a carrier. CONSTITUTION:Glycidyl monovinyl ester or glycidyl monovinyl ether (e.g., glycidyl methacrylate) being an (A)-component is crosslinked with alkylene glycol divinyl ester (e.g., ethylene glycol dimethacrylate) being a (B)-component to obtain a gel like copolymer based on (A)- and (B)-components. A bonding group, for example, an epoxy group, an amino group or a carboxyl group is introduced into this gel like copolymer and phenyl borate (e.g., aminophenyl borate) is bonded to said copolymer through the bonding group to obtain a porous copolymer. Because this porous copolymer is used as an adsorbing carrier for affinity chromatography, separation and purifying efficiency is enhanced and a working time can be shortened.

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, one of the conventional chromatography techniques, uses the affinity of pairs of substances that specifically interact with each other to separate and purify biological substances. It is useful in identifying and purifying a property, ie, a specific chemical structure on a molecule.

An adsorption carrier for affinity chromatography (affinity gel) is, for example, 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, phenylboric acid is known to easily and reversibly form a cyclic ester with a compound having a hydroxyl group on each adjacent carbon;
Nucleotides, catecholamines, carbohydrates and
-Affinity chromatography using phenylboric acid as a ligand is a useful means for separating and purifying many compounds belonging to RNA and the like.

For example, in separation, purification, and analysis by affinity chromatography in which phenylboric acid is used as a ligand and sugar 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, high porosity, easy binding of lycand and large immobilization capacity, chemical stability and sufficient stability under a wide range of pH ranges, salt concentrations, and temperatures. The characteristics include no volume change, sufficient mechanical strength and stability, good flow characteristics, and resistance 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.

The purpose of the present invention is to overcome the drawbacks of the conventional adsorption carriers for chromatography, and to provide a chromatography carrier that fully has 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

The present invention for solving the problem of chromatography provides an adsorption carrier for chromatography that can achieve the above object.

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 phenylboric acid 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 subjected to aqueous suspension polymerization in the presence of a water-insoluble organic diluent. It can be prepared by obtaining 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 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, as the component (B) which acts as a crosslinking component, an alkylene glycol having 2 or 3 carbon atoms or an ester of the polyalkylene glycol with acrylic acid or methacrylic acid is 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, polypropylene glycol dimethacrylate.

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 used may be one that is inert to the polymerization reaction, insoluble or sparingly soluble in water, but capable of dissolving the raw material monomer. 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. It is preferred that 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 preferably used in the present invention include cyclohexanone, chlorobenzene, benzene, toluene,
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 is not particularly limited, but 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 based on 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 with phenylboric acid. 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 the phenylboric acid include an epoxy group, a carboxyl group, and a formyl 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 the introduction of 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 10°C in water using sulfuric acid or hydrochloric acid as a catalyst.
This is done by heating to 0° C. for about 2 hours. The concentration of acid is preferably 0.1N to 0.5N. Ring-opening modification is performed almost quantitatively.

Examples of compounds having a glycidyl group that are bonded to the hydroxyl group generated by ring-opening modification include epihalohydrins such as epichlorohydrin and epibromohydrin, ethylene glycol diglycidyl ether, 1
．． Diglycidyl ethers such as 4-butanediol diglycidyl ether and polyethylene glycol diglycidyl ether, ■, 3-butadiene dieboxide, 1
, and alkyl diene dieboxides such as 7-octazidine dieboxide.

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 sodium hydroxide or potassium carbonate, or a hydrophilic copolymer is reacted with shrimp chlorohydrin. In an aqueous solution of an inorganic base such as sodium hydroxide or potassium carbonate containing sodium borohydride in the polymer, ethylene glycol or 1,4-butanediol diglycidyl ether,
Alternatively, it is carried out by reacting with polyethylene glycol diglycidyl ether. Repeating unit of polyethylene glycol diglycidyl ether used -02H
It is preferable to use one in which the number of 40- repeats is 1 to 10.

(ii) Introduction of a bonding group having a carboxyl group as a functional group For example, as a method for introducing a bonding group having a carboxyl group, an epoxy group based on component (A) of the gel copolymer according to the present invention or an epoxy group according to the present invention The epoxy group based on the glycidyl group of the modified gel-like copolymer is expressed by the general formula (1) (wherein R' represents a hydrogen atom, an amino group, an ω-aminoalkyl group, or an ω-carboxyalkyl group). on the epoxy group or glycidyl group, or when the amino compound in which R′ is other than the ω-carboxyalkyl group is reacted with an alkylenecarboxylic anhydride, Introducing a bonding group represented by the general formula (II) NHR (wherein R represents an ω-carboxyalkyl group, an ω-carboxyalkanoyl group, an ω-carboxyalkanoylamino group, or an ω-carboxyalkanoylaminoalkyl group). There is a way.

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

It represents the gel-like copolymer according to the present invention or the modified gel-like 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).

To the epoxy group of the gel-like copolymer according to the present invention or the glycidyl group of the modified gel-like copolymer according to the present invention, the reaction formula (D), general formula (D'), or The compound of general formula (F) can be reacted in the presence of a solvent, but if these compounds are liquid under the reaction conditions, no solvent may be used. 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 chlorobenzene, 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. Also, although it is not necessary to use a catalyst,
Alkali or alkaline earth metal hydroxides or carbonates such as sodium hydroxide and potassium carbonate can also be used.

Examples of the compound of general formula (D') include ethylenediamine,
1,3-diaminopropane, 1,4-diaminobutane,
■, 5-diaminopencune, 1,6-diamithexane, 1,7-diaminohbutane, 1,8-diaminooctane, 1,9-diaminononane, 1.10-diaminodecane, 1.12-diaminododecane Examples include diaminoalkanes such as.

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.

The weights 1t (a) and 2 (D) of the gel-like copolymer according to the present invention or the modified gel-like copolymer according to the present invention, the general formula (D'
) or the ratio of the weight (b) of the compound of general formula (F): a : b = 1 : 0.3 ~ IO More preferably, a : b = 1 : 0.3 ~ 3 Reaction degree 20 ~ 150℃, 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 it may be appropriately carried out by commonly used methods such as separate furnaces and washing.

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 no particular catalyst may be used, the pH of the reaction solution can be adjusted by adding an acid such as hydrochloric acid or sulfuric acid or an alkali such as sodium hydroxide or potassium carbonate.

In the reaction formula, the alkylenecarboxylic anhydride of general formula (E) or general formula (E') includes succinic anhydride, geltaric anhydride, adipic anhydride, pimelic anhydride, speric anhydride, and azelain. Examples include acid anhydrides and cepatic anhydride.

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

The weight (a') of the aminated product of the gel copolymer according to the present invention or the modified gel copolymer according to the present invention and the general formula (E)
Or the ratio of the weight (b') of the compound of general formula (E'): a':b'=l: 0.3-10, more preferably a':b'=1:0.3-3 Reaction temperature :0~150℃, more preferably room temperature~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 washing and washing may be carried out as appropriate.

Another method for introducing a bonding group having a carboxyl group as a functional group is to introduce an aminocarboxylic acid having a hydroxyl group into an epoxy group based on the component (A) of the gel copolymer according to the present invention or a modified method according to the present invention. There is a method of binding by acting on the glycidyl groups of a gel copolymer.

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

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 amino 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 the aminocarboxylic acid having a hydroxyl group 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 chlorobenzene, 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 restrictions on the reaction conditions, and for example, the same conditions as in the case of reacting the compound of general formula (1) above 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. .

When covalently bonding phenylboric acid to a binding group, aminophenylboric acid may be used; however, when covalently bonding phenylboric acid to a binding group, a catalyst or a reaction reagent may be used as appropriate depending on the functional group of the binding group. This can be carried out in an appropriate solvent using, for example, For example, as a catalyst,
Acids and alkalis such as hydrochloric acid, sodium carbonate or sodium bicarbonate are mainly used when the functional group is an epoxy group, and for example, N-hydroxysuccinimide, dicyclohexylcarbodiimide or 1-hydroxysuccinimide, dicyclohexylcarbodiimide or 1-
A condensing agent such as ethyl-3-(3-dimethylaminopropyl)carbodiimide is used when the functional group is a carboxyl group, a reducing agent such as sodium cyanoborohydride is used when the functional group is a formyl group, etc. can be exemplified. Water is usually used as a solvent, but if necessary, it can also be used as a phosphoric acid or acetate buffer, or an inorganic salt such as sodium chloride can be added thereto.

The conditions for the reaction with phenylboric acid 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 aminophenylboric acid: 1
:0.03-0.3, preferably 1:0.05-0.2
Reaction temperature: 0°C to room temperature, preferably 4°C to room temperature; Reaction time: 1 to 72 hours, preferably 2 to 12 hours.

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.

溌5Iυ Example: 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 phenylboric acid to this base through an appropriate bonding group. By using the adsorption carrier of the present invention, it can not only be used as a packing material for affinity chromatography similar to the conventional adsorption carrier using soft gel, but also particularly useful for pressure columns. The fact that it can be filled and used even under pressure has the greatest advantage of greatly shortening the working time and therefore greatly improving the efficiency of separation and purification. For example, it goes without saying that it is indispensable for applications in high-performance liquid chromatography and industrial separation and purification equipment.

In particular, while gel copolymers have many advantages as mentioned above, adsorption carriers to which phenylboric acid is covalently bonded via the bonding group are, for example, commonly used cyanogen bromide. Compared to adsorption carriers for affinity chromatography, etc., the adhesive does not fall off during use, and the length of the bonding group can be adjusted arbitrarily, so the adsorption ability of the ligand for the target substance is improved. In addition, the adsorption carrier for chromatography provided by the present invention has great effects in that it has excellent properties and has little nonspecific adsorption.

Tengu M Below, representative examples will be shown and more specifically explained about the method for producing the adsorption carrier for chromatography of the present invention from known substances. 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. To a suspension of 600 µm of gel (epoxy group: 0.3 mmol per 1 g of dry gel) in 4.5 mj2 of water was added 140 µm of 3-aminophenylborate hemisulfate, and the pH was adjusted to 8 with 4 molar aqueous sodium hydroxide solution. 5 and shaken at room temperature for 1 day. The gel was taken out of the oven and washed with a 1M aqueous sodium chloride solution and water. It was confirmed by titration with 0.1N sodium hydroxide that the adsorption carrier thus obtained supported 0.06 mmol of phenylboric acid per 1 g of dry gel.

Example 2 By carrying out the same operation as in Example 1 using shrimp chlorohydrin instead of 1,4-butanediol diglycidyl ether, 0.12 mmol of phenylboric acid was supported per 1 g of dry gel. An adsorption carrier was obtained.

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 the epoxy group was introduced with ethylene glycol diglycidyl ether. - After thoroughly washing 450 mg of gel into which a carboxyl group was introduced with aminobutyric acid with anhydrous dioxane, the mixture was added to 4 ml of anhydrous dioxane, and 80 μ of N-hydroxysuccinimide and 144 μ of dicyclohexylcarbodiimide were added thereto for 2 hours at room temperature. After shaking, the gel was taken out and quickly washed with 20 ml of anhydrous dioxane, 5 ml of methanol, and 3 ml of cold water in this order. This gel was added to a solution of 70 mg 3-aminophenylborate hemisulfate (7) 0.01 M sodium bicarbonate in 2 ml water and shaken overnight at room temperature. The gel was then taken out of the oven and washed with a 1 molar aqueous sodium chloride solution and water. It was confirmed that the adsorption carrier thus obtained supported 0.03 mmol of phenylboric acid per 1 g of dry gel.

Example 4 To 1.0 g of an epoxy group-containing gel copolymer obtained from glycidyl methacrylate and ethylene glycol dimethacrylate, 4 tell of concentrated ammonia water was added, and the mixture was shaken at 40°C for 5 hours. The gel was taken out of the oven, washed with water and dried, then added to a solution of 1.0 g of succinic anhydride in 5 ml of 0.1N sodium chloride water, the pH was adjusted to 6, and the gel was shaken at room temperature for one day. Remove the gel from the furnace and add 0, 1N sodium bicarbonate, 0
．． It was washed successively with 1N hydrochloric acid and water and dried. This gel contains 0.00 carboxyl groups per Ig (dry). It was confirmed by titration with 0 and 1 N sodium hydroxide that it contained 100 mmol. Next, 600 µm of this gel was added to 10 m/ml of 0.1 mol sodium chloride, and while keeping the temperature at 0°C, 164 µm of 3-aminophenylboric acid hemisulfate was added to adjust the pH to 7.0, and then 1-ethyl-3- 165 ml of (3-dimethylaminopropyl)carbodiimide hydrochloride was added and the mixture was shaken for 3 hours. After standing at 4°C overnight, the gel was taken out and washed with 0.1N sodium chloride. It was confirmed by measuring unreacted 3-aminophenylboric acid using an absorbance method that the adsorption carrier thus obtained supported 0.47 mmol of phenylboric acid per gram of dry gel.

Example 5 An adsorption carrier carrying 0.32 mmol of phenylboric acid per 1 g of dry gel was obtained by carrying out the same operation as in Example 4 using 1,3-diaminopropane instead of concentrated ammonia water. It was done.

Test Examples The adsorption carriers obtained in Examples 1 to 5 were packed into a stainless steel column with a diameter of 4.6 fi and a length of 75 fins, and sample compounds were analyzed using a high performance liquid chromatography device.
The results shown in Table 1 were obtained.

Analysis conditions: Eluent: 0.1M potassium phosphate buffer (
pH 7.5), elution rate: 1. Om7! /min
, Detector: ultraviolet spectrophotometer (260 nm) or differential refractometer.

Table 1 Sample retention time)

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 phenylboric acid covalently bonded to a group through a bonding group.