JPS62153754A - Adsorber 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 iminodiacetic acid 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 are main components and the (A)-component is crosslinked with the (B)-component, iminodiacetic acid 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 Field of Industrial Application The present invention relates to an adsorption carrier for chromatography that can be used in various chromatography including affinity chromatography.

As a type of chromatography technology, affinity chromatography is a method of separating and purifying biological substances by utilizing the affinity of pairs of substances that specifically interact with each other. It is useful for identifying and purifying biological properties, ie, certain chemical structures on molecules.

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, iminodiacetic acid is known to form chelates with many metals such as cobalt, nickel, copper, zinc, and cadmium as a royal ligand due to the two oxygens and one nitrogen in the molecule. On the other hand, it is known that proteins also coordinate with metals depending on the type of their constituent amino acids, and therefore, an adsorption carrier for chromatography using iminodiacetic acid as a ligand is useful for separating and purifying proteins in a metal-mediated state. It is considered.

For example, in separation, purification, and analysis by affinity chromatography in which iminodiacetic acid is used as a ligand and protein 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,
These include being sufficiently stable and not changing in volume under a wide range of salt concentration and temperature conditions, having sufficient mechanical strength and stability, and having good flow characteristics, and being 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.

The object of the present invention is to overcome the drawbacks of the conventional adsorption carriers for chromatography, and to achieve 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.

The present invention 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 iminodiacetic acid covalently bonded to an epoxy group based on the component (A) of the gel-like copolymer according to the present invention via a bonding group. This 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 aqueous suspension polymerized in the presence of a water-insoluble diluent). 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.

As the component (A), glycidyl ester of monovinyl carboxylic acid having 3 to 12 carbon atoms or UIB3 to 12
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 3 alkylene glycol or an ester of the polyalkylene glycol and 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 and 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%. Less than half of the component (A) is a comonomer, namely methyl methacrylate, methyl agerylate,
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 content 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. The amount of epoxy oxygen expressed as epoxy oxygen in the resulting polymer is preferably greater than 1% by weight, up to about 10% by weight94.

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, n-propyl acetate, n-butyl acetate, n-butyl acetate,
-Octane etc.

The aqueous suspension polymerization can be carried out in the presence of an aM group-based 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 iminodiacetic 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 iminodiacetic acid include an epoxy group and a carboxyl 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 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.

Compounds having a glycidyl group that are bonded to the hydroxyl group produced by ring-opening modification include shrimp halohydrins such as shrimp chlorohydrin and epibromohydrin, ethylene glycol diglycidyl ether, 1
．． Diglycidyl ethers such as 4-butanediol diglycidyl ether and polyethylene glycol diglycidyl ether, 1.3-butadiene dieboxide, 1
, and alkyl diene dieboxides such as 7-octazidine dieboxide.

In order to react with a compound containing a glycidyl group, for example, a ring-opening modified gel copolymer is reacted with epichlorohydrin in an aqueous solution of an inorganic base such as sodium hydroxide or potassium carbonate, or a hydrophilic copolymer is reacted with epichlorohydrin. 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. 1 unit of polyethylene glycol diglycidyl ether used -〇
It is preferable to use one having a repeating number of 2 H40- of 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 may be used. 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 (n) 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.

In the following reaction formula, (base+CH-CH2 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 the base + CH-CH2 in these copolymers. represents one epoxy group or glycidyl group, where m is 1 or 2, n, p, q and r are each 1
or more (more preferably 1 to 12).

-'''-Self/Main Chaki 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 is combined with '] 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.As a solvent, , water is usually used, but other hydrocarbons such as n-hexane, benzene, and xylene, ethers such as tetrahydrofuran and dioxane, chlorinated hydrocarbons such as chloroform and chlorohenzene, alcohols such as ethanol and methyl cellosolve, In addition to ketones such as acetone and methyl isobutyl ketone, dimethylformamide and dimethyl sulfoxide are also used, and these may be used alone or in a mixed solvent system.Also, no catalyst may be used, but
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,
1.5-diaminopentane, 1.6-diamithexane, 1.7-diaminohbutane, 1,8-diaminooctane, 1.9-diaminononane, 1.10-diaminodecane, 1,12-diaminododecane, etc. diaminoalkanes.

-I Examples of the compound of 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=1: 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 10atII+, 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.

The 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'=1:0.3-10, more preferably a':b'=l:Q, 3-3 Reaction temperature -0 to 150°C, more preferably room temperature to 100°C
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.

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-
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 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 limitations on the reaction conditions, and for example, the same conditions as in the case where the compound of general formula (I) 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.

When covalently bonding iminodiacetic acid to a bonding 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 bonding group. For example, as a catalyst, acids such as hydrochloric acid, sodium carbonate or sodium bicarbonate, or alkali are mainly used when the functional group is an epoxy group, and as a reaction reagent, for example, N-hydroxysuccinimide, dicyclohexylcarbodiimide or A condensing agent such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide is used when the functional group is a carboxyl group, and a reducing agent such as sodium cyanoborohydride is used when the functional group is a formyl group. Examples include: Water is usually used as a solvent, but if necessary, a phosphoric acid or acetate buffer may be used, or an inorganic salt such as sodium chloride may be added.

The reaction conditions with iminodiacetic 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 bonding group is attached and iminodiacetic acid: 1:0.0
3-0.3, preferably 1:0.05-0.2; 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, and conventional methods such as separation and washing may be carried out as appropriate.

The adsorption carrier for chromatography provided by the present invention uses a base that has all the properties desired as an adsorption carrier for affinity chromatography as described above, and binds it to the base through an appropriate bonding group. Since iminodiacetic acid is 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 conventional adsorption carriers using soft gels, but it can also be packed into pressure-resistant columns (pressurized), which means that the work time can be reduced. Since the width can be shortened to a large extent, the biggest advantage is that the efficiency of separation and purification can be greatly improved. Needless to say, it is essential for measuring applications.

In particular, compared to gel-like copolymers which have many advantages as described above, adsorption carriers to which iminodiacetic acid is bonded via bonding groups (7) and covalent bonds as described above, for example, 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, as 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 described below 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 group: 0.3 mmol per 1 g of dry gel) was mixed with 0.53 g of iminodiacetic acid disodium salt.
2M aqueous solution of sodium carbonate'/f! After adding to the solution in L3mp and shaking at 65°C for 24 hours, it was left at 4°C overnight. The gel was renewed and washed with 10% aqueous acetic acid and water. It was confirmed by titrating the adsorption carrier with 0.1N sodium hydroxide solution that the adsorption carrier thus obtained supported 0.090 mmol of iminodiacetic acid per 1 g of dry gel.

Example 2 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 diglyndyl ether. - After thoroughly washing 1.0 g of gel into which carboxyl groups were introduced with aminobutyric acid with anhydrous dioxane, it was added to 4 ml of anhydrous dioxane, and then 80 μg of N-hydroxysuccinimide and 1 ml of dicyclohexylcarbodiimide were added.
After adding 44■ and shaking at room temperature for 2 hours, the gel was renewed.
20ml of anhydrous dioxane, 6ml of methanol, 3ml of cold water
It was quickly washed in the order of ml. This gel was prepared by mixing 0.2 g of iminodiacetic acid disodium salt with 0.1 molar sodium bicarbonate in 2 ml of water. After shaking at room temperature for 2 hours, the mixture was left at 4°C overnight.

7 units of gel and 1M sodium chloride water? Washed with liquid 8 and water. It was confirmed that the adsorption carrier thus obtained supported 0.052 mmol of iminodiacetic acid per gram of dry gel.

Test Example 1 The adsorption carrier obtained in Example 1 had a diameter of 4.6 tm and a length of 7
Adenosine-5'-monophosphate (AMP), guanosine-5'-monophosphate (GMP), and cytidine-5'-monophosphate were packed in a 5 mm stainless steel column using a high performance liquid chromatography device. When (CMP) was analyzed, a chromatogram as shown in FIG. 1 was obtained. Analysis conditions: Eluent: 0.05 molar ethylmorpholine acetate buffer (pH 6,0). Elution rate: 1. Omβ/min, detector: ultraviolet spectrophotometer 28
0nm.

Test Example 2 In Test Example 1, after flowing a 50 mmol copper sulfate aqueous solution through the column to coordinate copper, the column was thoroughly washed with the eluent used, and analysis was conducted in the same manner as in Figure 2. A romatogram was obtained.

Test Example 3 Analysis was conducted in the same manner as in Test Example 2 except that uridine, cytidine, guanosine, and adenosine were used instead of AMP, GMP, and CMP, and a chromatogram as shown in FIG. 3 was obtained.

31... uridine and cytidine, 32...Guanosine, 33...Adenosine.

Test Example 4 In Test Example 1, 0.5 mol of sodium chloride was added to 0.1 mol sodium acetate buffer (pH 7.7) instead of 0.05 mol ethylmorpholine acetate buffer as the eluent ( AMP, GMP
When the analysis was carried out in the same manner except that bovine serum albumin (BSA) and transferrin were used in place of CMP, a chromatogram as shown in FIG. 4 was obtained.

Test Example 5 In Test Example 2, 0.15 molar ammonium chloride was added to eluent ■ and 0.05 molar Tris-HCl buffer (pH 8,0) instead of 0.05 molar ethylmorpholine acetate buffer as the eluent. (hereinafter referred to as eluent ■),
Analysis was carried out in the same manner except that BSA was used instead of MP, GMP and CMP, and a chromatogram as shown in FIG. 5 was obtained.

Test Example 6 In Test Example 2, zinc sulfate was used in the column instead of the copper sulfate aqueous solution, and eluent ■ and eluent ■ were used instead of the 0.05 M ethylmorpholine acetate buffer as the eluent.
, BSA and transferrin were used in place of GMP and CMP, and the analysis was carried out in the same manner, and a chromatogram as shown in FIG. 6 was obtained.

Figures 1 and 2 show that the elution of AMP and GMP is delayed when the ligand is coordinated with copper. Third
The figure also shows that the elution of guanosine and adenosine is delayed due to copper coordination.

Figure 4 shows that BSA and transferrin do not adsorb unless copper is coordinated, but BSA is adsorbed with eluent ■ in the presence of copper, and when switching to eluent ■ (arrow in the figure)
This shows that the sample elutes (Figure 5). Figure 6 shows the case where zinc is used as the coordinating metal, and with the eluent ■, BSA is not adsorbed but transferrin is adsorbed, and this is due to the eluent ■ (
(arrow in the figure) indicates elution.

[Brief explanation of drawings]

1 to 6 are chromatograms showing patterns of separation, adsorption, or elution by a high performance liquid chromatography column using the chromatography adsorption carrier of the present invention, respectively. Agent Patent Attorney Jo Taira Yamashita Figure 1 Figure 2 Figure 3 List

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). 1. An adsorption carrier for chromatography, comprising a porous copolymer containing iminodiacetic acid bound to a group by a covalent bond via a bonding group.