JPH11322997A - Preparation of porous bead - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable wide control of the kind and introduced amount of functional groups on porous beads by discharging a uniform polymerizable solution which is a mixture of a crosslinked polymer-generating material, a polymerizable compound having a functional group and a phase separator as an oil drop and irradiating an active energy ray to the same. SOLUTION: (A) A crosslinked polymer-generating materials comprising a polymerizable compound having two or more unsaturated groups capable of polymerizing by irradiation of an active energy ray, (B) a polymerizable compound having a functional group selected from ion groups, carboxylic group, phosphoric acid residue, amino group, sugar residues, amino acid residues and an unsaturated group capable of polymerizing by irradiation of the active energy ray and (C) a phase separator which is compatible with or soluble in component A or soluble in a uniform solution of component A and a solvent and which is not compatible with the crosslinked polymer generated by irradiating the active energy ray to component A and which is inert to the active energy ray are used. After the irradiation of the active energy ray, the phase separator and unreacted polymerizable solution are removed.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a protein, a food, an electronic industry, a wastewater treatment, an artificial organ, a medical treatment, etc.
Removal and recovery of colloids, bacteria, etc., adsorption removal of harmful gases, decolorization, collection and removal of harmful substances in water, collection of heavy metals, carriers for immobilization of proteins, enzymes, bacteria, etc., carriers for affinity chromatography, etc. The present invention relates to a method for producing porous beads used for the purpose.

[0002]

2. Description of the Related Art Japanese Patent Application Laid-Open No. Sho 50-12176, Journal of Polymer Science Part A-1, Vol.
689, 1968 (Journal of Pol)
ymer Science: Part A-1, Vo
l. 62689 (1968)) has proposed an oil-in-water emulsion polymerization method. Further, as a method for producing porous particles having an average particle diameter of one hundred to several hundred μm, Japanese Patent Application Laid-Open
Japanese Patent Application No. 78541 proposes a method in which a polymer solution is ejected into a gas phase as droplets of a uniform size, and then the polymer solution is allowed to penetrate into a coagulating liquid to be coagulated.

[0003]

However, in the former method, there is a restriction that the solvent of the polymerizable monomer must be less than a certain solubility in water, and a stable condition is required if oil droplets in water are not smaller than a certain size. It is difficult to produce porous particles having a particle size of several hundred μm to several millimeters due to the restriction that a suitable emulsion cannot be obtained. In addition, it is difficult to freely adjust the particle diameter of the particles and the pore diameter of the porous portion by this method. Furthermore, solvents such as benzene, carbon tetrachloride, and chloroform used in this method have a drawback that they are harmful substances that may cause health problems.

In the latter method, it is possible to control the particle size and particle size distribution of the particles. However, since the method does not have any functional element, it is difficult to introduce a new functional functional group. In addition, it is necessary to use a method such as surface treatment for the porous particles, and there is a disadvantage that the number of treatment steps increases and the production cost increases.

The present inventors discharge a uniform polymerizable solution comprising a polymerizable compound having a polymerizable unsaturated group and a phase separator by irradiation with active energy rays in the form of droplets, and apply the active energy rays thereto. A novel method for producing porous beads characterized by irradiation (JP-A-6-248107) was proposed. This method is characterized in that the particle size and the pore size of the porous portion can be easily controlled, and that no harmful solvents such as benzene, carbon tetrachloride, and chloroform are used unlike the emulsion polymerization method.

However, the above-mentioned invention (Japanese Patent Laid-Open No. 6-2481)
No. 07) did not mention the introduction of a functional functional group into the porous beads and the development of a new function associated therewith.

The problem to be solved by the present invention is that the type and amount of functional functional groups to be introduced can be widely adjusted, several types of functional functional groups can be simultaneously introduced into beads, the particle size can be easily controlled, and productivity can be improved. To provide a method for producing a porous bead having a high density.

[0008]

Means for Solving the Problems The inventors of the present invention have conducted intensive studies to solve the above-mentioned problems, and as a result, the type and the amount of the functional functional groups to be introduced can be controlled widely, and the obtained porous The discovery of a new function in the beads led to the completion of the present invention.

That is, in order to solve the above-mentioned problems, the present invention provides (1) a crosslinked polymer-forming material comprising a polymerizable compound having two or more unsaturated groups per molecule which can be polymerized by irradiation with active energy rays ( A) and a polymer having a functional functional group selected from the group consisting of an ionic group, a carboxyl group, a phosphoric acid residue, an amino group, a sugar residue and an amino acid residue, and an unsaturated group polymerizable by irradiation with active energy rays. Soluble or soluble in the water-soluble compound (B) and the crosslinked polymer-forming material (A), or dissolved in a homogeneous solution of the crosslinked polymer-forming material (A) and a solvent to form a crosslinked polymer. Uniform polymerizability obtained by mixing a phase separation agent (C) which is incompatible with the crosslinked polymer produced by irradiating the material (A) with active energy rays and which is inactive with respect to active energy rays Solution in droplet form Discharged was, this was irradiated with an active energy ray, then, to provide a method of manufacturing a porous beads and removing the polymerization solution phase separation agent (C) and unreacted.

Further, the present invention is characterized in that (2) the method of ejecting the liquid in the form of droplets is a method of extruding the polymerizable solution from the nozzle under the condition that the polymerizable solution becomes droplets during free fall.
The manufacturing method according to the above (1) is provided.

Further, the present invention is characterized in that (1) 1 to 5 parts by weight of a phase separator (C) is used per 1 part by weight of the crosslinked polymer forming material (A). Or, the production method according to (2) is provided.

Further, according to the present invention, (4) the polymerizable compound (B) may have a (meth) acrylate having an ionic group, a (meth) acrylate having a carboxyl group, a phosphoric acid (meth) acrylate, or a (meth) acrylate having an amino group. ) Acrylate, sugar (meth) acrylate and amino acid (meth)
(1) the (meth) acrylic polymerizable compound selected from the group consisting of acrylates,
(2) The method according to (3) is provided.

Further, the present invention provides (5) 1 part by weight of the crosslinked polymer-forming material (A) and 0.1 parts by weight of the polymerizable compound (B).
The above (1), characterized in that it is used in an amount of from 03 to 3 parts by weight.
(2) The method according to (3) or (4) is provided.

[0014]

Next, the present invention will be described in detail. The crosslinked polymer forming material (A) used in the present invention is:
1 unsaturated group polymerizable by irradiation with active energy rays
It contains a polymerizable compound having two or more in the molecule (hereinafter, referred to as polymerizable compound (A)) as an essential component. The polymerizable compound (A) is not particularly limited as long as it is a substance that can be polymerized by irradiation with active energy rays, regardless of whether it is organic or inorganic, as long as it is a radical polymerizable compound, an anionic polymerizable compound, a cationic polymerizable compound, and the like. May be. For example, a polymerizable compound having a vinyl group, a vinylidene group, an acryl group, a methacryl group (the acryl group and the methacryl group are collectively referred to as a (meth) acryl group, and the same applies to a (meth) acrylate). Those having a high polymerization rate by irradiation with energy rays are preferred.

Specific examples of the polymerizable compound (A) used in the present invention include diethylene glycol di (meth) acrylate, neopentyl glycol di (meth) acrylate, 1,6-hexanediol di (meth) acrylate, and polyethylene glycol. Di (meth) acrylate,
Polymerizable compounds having two unsaturated groups, such as 2,2'-bis {4- (meth) acryloyloxypolyethyleneoxyphenyl} propane and dicyclopentenyldi (meth) acrylate, trimethylolpropane tri (meth) acrylate A polymerizable compound having three unsaturated groups, such as
In addition to a polymerizable compound having four unsaturated groups, such as pentaerythritol tetra (meth) acrylate, a polymerizable compound having six unsaturated groups, such as dipentaerythritol hexa (meth) acrylate, bisphenol A-diepoxy- (Meth) acrylic acid adducts, (meth) acrylic acid esters of epoxy resins, (meth) acrylic acid esters of polyether resins, (meth) acrylic acid esters of polybutadiene resins, having (meth) acrylic groups at molecular terminals Polymerizable compounds having a weight average molecular weight of 500 to 50,000, such as a polyurethane resin, are exemplified. These compounds (A) may be used alone or in combination of two or more.

Further, in the crosslinked polymer forming material (A), a polymerizable compound having one unsaturated group in one molecule which can be polymerized by irradiation with active energy rays can be used in combination. By using them together, the crosslink density, pressure resistance, heat resistance, solvent resistance and the like of the obtained porous beads can be adjusted.

The polymerizable compound having one unsaturated group in one molecule is not particularly limited as long as it is a substance which is polymerized by irradiation with active energy rays to be a polymer. Specific examples thereof include, for example, ethyl (meth) Acrylate, hydroxy (meth) acrylate, n-butyl (meth) acrylate, hexyl (meth) acrylate, phenyl (meth)
Acrylate, phenyl cellosolve (meth) acrylate, n-vinylpyrrolidone, isobornyl (meth) acrylate, dicyclopentenyl (meth) acrylate,
Examples thereof include lauryl (meth) acrylate, 2-methoxyethyl (meth) acrylate, 2- (meth) acryloyloxyethyl hydrophthalate, and trifluoroethyl (meth) acrylate.

The polymerizable compound (B) used in the present invention
Is an ionic group, a carboxyl group, a phosphoric acid residue, an amino group,
It is not particularly limited as long as it has a functional functional group selected from the group consisting of a sugar residue and an amino acid residue and an unsaturated group polymerizable by irradiation with active energy rays. And a compound having a (meth) acryl group.

Specific examples of the polymerizable compound (B) include, for example, (meth) acrylate having an anion such as sodium sulfonic acid (meth) acrylate, sodium 2-methylpropane-2-acrylamide sulfonate, and tri (meth) acrylate. (Meth) acrylate having a cation such as -butyl {2- (meth) acryloyloxyethyl} phosphonium chloride, dimethylaminoethyl (meth) acrylate quaternary compound, 2- (meth) acryloyloxypropyl hydrogen phthalate, 2-acryloyl (Meth) acrylate having a carboxyl group such as oxyethyl succinic acid, (meth) acrylic acid, 2- (meth) acryloyloxypropylhexahydrohydrogenphthalate, mono {2- (meth) acryloyloxyethyl} acid phosphate , 10- (meth) acryloyloxydecyl dihydrogen phosphate, beta-
Phosphoric acid (meth) acrylate such as (meth) acryloyloxyethylphenylphosphoric acid, (meth) acrylamide, N, N-dimethyl (meth) acrylamide, N-isopropyl (meth) acrylamide, N, N-dimethylaminopropyl (meth) And (meth) acrylates having an amino group such as acrylamide and N-methylenebis (meth) acrylamide, sugar (meth) acrylate and amino acid (meth) acrylate.

In the present invention, "sugar (meth) acrylate"
Means that a sugar residue is covalently bonded in the molecule of the polymerizable compound (B). Examples of the sugar residue include monosaccharides such as glucose, galactose, and mannose and derivatives of these monosaccharides (eg, methylglucoside), and disaccharides such as maltose (maltose), cellobiose, lactose (lactose), and sucrose. Derivatives thereof, oligosaccharides such as cyclodextrin or derivatives thereof, and polysaccharides such as starch are exemplified.

Specific examples of the sugar acrylate include, for example, 6- (meth) acryloyl (1-O-) n-butylglucoside, 6- (meth) acryloylglucose,
-(Meth) acryloyl cellobiose, 6,6′-bis (meth) acryloyl cellobiose and the like.

In the present invention, “(meth) of amino acid”
“Acrylate” indicates that an amino acid residue is covalently bonded in the molecule of the polymerizable compound (B). Examples of the amino acid residue include tryptophan, alanine, isoleucine, methionine, phenylalanine, proline, valine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, histidine, hydroxylysine, hydroxyproline, lysine, and serine. Examples include amino acids and their derivatives such as threonine tyrosine, peptides such as glycylglycine and glycylalanylphenylalanine, and proteins such as polypeptides and albumin. Specific examples of the amino acid (meth) acrylate include, for example, N
α-{(meth) acryloyloxyethylaminocarbonyl} lysine, Nα-{(meth) acryloyloxyethylaminocarbonyl} phenylalanine and the like. These compounds (B) may be used alone or in combination of two or more.

The use of the polymerizable compound (B) having a functional functional group allows the porous beads to have an ion exchange property,
Functionality such as low protein adsorption, biocompatibility, selective adsorption to proteins, toxins, enzymes, etc., and affinity properties can be imparted. Further, these compounds (B)
By combining a plurality of them, changing the ratio thereof, or further introducing another functional group (for example, a sulfonic acid group) into the introduced functional functional group (for example, a sugar residue), thereby improving the antithrombotic or optical properties. New functions such as activity and physiological activity can be imparted.

With respect to the amount of the polymerizable compound (B) used,
0.03 to 1 part by weight of the crosslinked polymer forming material (A)
A range of 3 parts by weight is preferable, and 0.1 to 1 part by weight is particularly preferable. If the amount is less than 0.03 part, the function of the functional functional group is hardly exhibited, and if the amount exceeds 3 parts, beads having a high hydrophilicity are easily swelled when a functional group having a high hydrophilicity is used, resulting in poor strength. This is not preferable because it becomes sufficient or the width of adjustment of the hole diameter becomes narrow.

The phase separating agent (C) used in the present invention is compatible with or dissolved in the crosslinked polymer forming material (A),
Alternatively, it is dissolved in a homogeneous solution of the crosslinked polymer forming material (A) and a solvent, and is not compatible with the crosslinked polymer formed by irradiating the crosslinked polymer forming material (A) with active energy rays. There is no particular limitation as long as it is inert to active energy rays. Specifically, for example, dialkyl ketones such as diisobutyl ketone, fatty acid alkyl esters such as methyl caprate, polyethylene glycol monoester, polyethylene glycol monoether, glycerin mono, di and triester, alcohols and water A mixed solution, a mixture of a water-soluble polymer such as polyvinylpyrrolidone and polyethylene glycol and a polar solvent such as dimethylacetamide, and the like can be given. These phase separation agents (C) can be used alone or in combination of two or more.

The phase-separating agent (C) is prepared by the following methods: the method of producing beads, the types of the polymerizable compounds (A) and (B) used, the required viscosity of the polymerizable solution, and the solubility of additives. ,
It can be appropriately selected depending on the pore diameter, pore shape, and the like required for the beads.

The amount of the phase separator (C) used is preferably in the range of 1 to 5 parts by weight, more preferably 2 to 3.5 parts by weight, per 1 part by weight of the crosslinked polymer forming material (A). If the amount is less than 1 part, the porosity of the pores becomes low, and if the amount exceeds 5 parts, the strength of the beads becomes insufficient, which is not preferable.

By selecting a crosslinked polymer-forming material (A), a polymerizable compound (B) having a functional group and an unsaturated group polymerizable by irradiation with active energy rays, a phase separator (C), etc. Beads having excellent pore size, strength, flexibility, heat resistance, chemical resistance, swelling resistance, and functionality can be produced.

A polymer solution, that is, a crosslinked polymer forming material (A) and a polymerizable compound (B) having a functional functional group
If necessary, other additives such as a polymerization initiator, a viscosity modifier, an antifungal agent, and an indicator may be added to the mixed solution of the phase separation agent (C).

In the method for producing a porous bead of the present invention, the polymerizable solution is extruded from a nozzle under the condition that the polymerizable solution is formed into a droplet or a droplet during free fall, and the droplet is irradiated with an active energy ray. The manufacturing method is characterized in that: It is also possible that the extrusion from the nozzle is a spray. As the polymerizable solution, for example, the polymerizable solution may be dropped naturally from a glass pipette, or may be sent using a liquid sending pump.

The particle size of the porous beads can be controlled by adjusting the size of the cross-sectional area of the discharge port of the nozzle and the linear velocity of the discharge, or by controlling the viscosity of the polymerizable solution.

The active energy rays used in the production method of the present invention include electron beams, gamma rays, X-rays, ultraviolet rays, visible rays and the like. Above all, ultraviolet light is preferable from the viewpoint of simplicity of equipment and handling. The intensity of the irradiated ultraviolet light is preferably from 10 to 5000 mW / cm ² , and the irradiation time is preferably from about 0.01 to 10 seconds. In this case, it is preferable to add an ultraviolet polymerization initiator to the polymerizable solution for the purpose of increasing the polymerization rate.

The ultraviolet polymerization initiator used in the present invention is not particularly limited, but is preferably one that can be dissolved in a polymerizable solution. Specifically, for example, p-ter
t-butyltrichloroacetophenone, 2,2'-diethoxyacetophenone, 2-hydroxy-2-methyl-
Acetophenones such as 1-phenylpropan-1-one and hydroxycyclohexylphenyl ketone, benzophenone, 4,4'-bisdimethylaminobenzophenone, 2-chlorothioxanthone, 2-methylthioxanthone, 2-ethylthioxanthone, 2-isopropylthioxanthone and the like Ketones, benzoin, benzoin methyl ether, benzoin isopropyl ether, benzoin ethers such as benzoin isobutyl ether,
And benzyl ketals such as benzyl dimethyl ketal.

The obtained beads may be washed with a phase separator (C), an unreacted polymerizable compound (A) or (B), an unreacted polymerizable compound (A) or (B), an ultraviolet polymerization initiator, and other components. Remove additives. The detergent may be any as long as it can dissolve the phase separation agent (C), the polymerization initiator, unreacted components and other additives. Further, it is preferable to use alcohols such as ethanol or water from the viewpoint of avoiding the influence on the environment and the human body. When water or a water-soluble substance is used as the phase separation agent (C), it can be used without washing.

The porous beads produced according to the present invention have a capacity of 0.1%.
It has a large number of communication holes of from 01 μm to 10 μm, and the particle size is from 0.01 to 7 mm. Porous beads having a small particle size can be obtained by spraying a polymerizable solution from a nozzle and irradiating with an active energy ray.

The shape of the porous beads produced in the present invention is not necessarily spherical, but may be particles of any shape such as spheroid, raindrop, disk, dish, hemisphere and the like.

[0037]

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to Examples, but the scope of the present invention is not limited thereto.

[Definition of Measurement Items] Measurement was carried out by the following method. << Elemental Analysis >> The elemental component of the polymer constituting the beads is X
Measured by photoelectron spectroscopy (ESCA) with line excitation,
The element composition was represented by a composition ratio of the number of atoms. The apparatus used was an X-ray photoelectron analyzer ESCA850, manufactured by Shimadzu Corporation.

Example 1 <Preparation of Polymerizable Solution> As the polymerizable compound (A), a urethane acrylate having a weight average molecular weight of 2,000 and three acryl groups on average per molecule (Dainippon Ink Chemical Industry Co., Ltd.) 76 parts Unidick V-4263 manufactured by K.K., hexanediol diacrylate (Daiichi Kogyo Seiyaku Co., Ltd.)
19 parts of New Frontier HDDA), 3 parts of mono (2-acryloyloxyethyl) acid phosphate (light ester PA manufactured by Kyoeisha Chemical Co., Ltd.) as a polymerizable compound (B) having a functional functional group, photopolymerization As an initiator, Irgacure 184 (manufactured by Ciba Geigy)
2 parts, as phase separator (C), methyl caprate 180
Parts (manufactured by Wako Pure Chemical Industries, Ltd.) were uniformly mixed to obtain a polymerizable solution (1).

<Preparation of porous beads> Inner diameter 0.53 mm
Of the polymerizable solution (1) using a needle nozzle of
When the polymerizable solution was extruded into a nitrogen atmosphere at a rate of ml / min (discharge linear velocity of 151 cm / sec), the polymerizable solution had a columnar shape immediately below the nozzle, and turned into a droplet at about 10 cm or less below the nozzle. A range of 30 to 60 cm below the nozzle was irradiated with ultraviolet rays having a wavelength of 365 nm using a condenser mirror by a 6 KW metal halide lamp.

The cured beads were dropped into a container at 50 cm below the nozzle and containing a 50% aqueous ethanol solution. The obtained beads were further washed with ethanol and water, and dried under reduced pressure to obtain white beads having no gloss on the surface.

<Characteristics of beads> The beads have a diameter of 0.9.
When observed with a scanning electron microscope, the outer surface had a mesh-like structure with a pore diameter of about 1 μm, and the cut inside had a porous structure in which polymer fine particles were bonded to each other. The gap between the fine particles was about 0.8 μm.

The element ratio on the surface of the beads is as follows: carbon (C): oxygen (O): nitrogen (N): phosphorus (P) = 71.
6: 24.8: 3.4: 0.2. That is, phosphorus (P) derived from a phosphate group was detected on the bead surface.

When 1.0 g of the beads was added to 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 19.4 mg of albumin was adsorbed on the beads. did.

Example 2 <Preparation of Polymerizable Solution> As the polymerizable compound (A), a urethane acrylate having a weight average molecular weight of 2,000 and an average of three acrylic groups in one molecule (Dainippon Ink Chemical Industry) 56 parts of Unidick V-4263), hexanediol diacrylate (Daiichi Kogyo Seiyaku Co., Ltd.)
14 parts of New Frontier HDDA), 30 parts of mono (2-acryloyloxyethyl) acid phosphate (Light Ester PA, manufactured by Kyoeisha Chemical Co., Ltd.) as a polymerizable compound (B) having a functional functional group, photopolymerization 2 parts of Irgacure 184 (manufactured by Ciba Geigy) as an initiator, and methyl caprate 1 as a phase separator (C)
80 parts (manufactured by Wako Pure Chemical Industries, Ltd.) were uniformly mixed to obtain a polymerizable solution (2).

<Preparation of Porous Beads> In the same manner as in Example 1, white beads having no gloss on the surface were obtained.

<Characteristics of beads> The beads had a diameter of 0.9.
When observed with a scanning electron microscope, the outer surface had a mesh-like structure with a pore diameter of about 1 μm, and the cut inside had a porous structure in which polymer fine particles were adhered to each other. The gap between the fine particles was about 0.8 μm.

The element ratio on the surface of the beads is C: O:
N: P = 64: 31: 2: 3. That is, phosphorus (P) derived from a phosphate group was detected on the bead surface.

When 1.0 g of the beads was put in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 0.3 mg of albumin was adsorbed on the beads. did.

A similar adsorption experiment was performed using a 0.3% bovine serum albumin aqueous solution (pH 3.5), and it was found that 0.65 mg of albumin was adsorbed on the beads.

As described above, the results of Examples 1 and 2 show that as the amount of phosphate groups introduced into the porous beads increases, the hydrophilicity of the beads increases and the hydrophobic interaction with the protein decreases. did. From this, it became clear that the amount of adsorbed protein, which is one of the indicators of biocompatibility, can be controlled by the amount of phosphate groups introduced.

Example 3 <Preparation of Polymerizable Solution> As the polymerizable compound (A), a urethane acrylate having a weight average molecular weight of 2,000 and an average of three acrylic groups per molecule (Dainippon Ink Chemical Industry) (Manufactured by Unidick V-4263) 77.6
Part, ethylene oxide-modified bisphenol A diacrylate (New Frontier B, manufactured by Daiichi Kogyo Seiyaku Co., Ltd.)
PE-4) 19.4 parts, as a polymerizable compound (B) having a functional functional group, sodium sulfonate methacrylate (Nippon Emulsifier Co., Ltd., Antox MS-2N)
3 parts, 2 parts of Irgacure 184 (manufactured by Ciba Geigy) as a photopolymerization initiator, and 100 parts of 2-propanol (manufactured by Wako Pure Chemical Industries, Ltd.) and 40 parts of distilled water as a phase separator (C). The mixed solution is uniformly mixed, and the polymerizable solution (3)
I got

<Preparation of Porous Beads> White beads having no gloss on the surface were obtained in the same manner as in Example 1.

<Characteristics of beads> The beads had a diameter of 0.9.
When observed with a scanning electron microscope, both the outer surface and the cut interior had a porous structure in which polymer fine particles adhered to each other. The gap between the particles is about 2μm
Met.

The element ratio on the surface of the beads is C: O:
N: Sulfur (S) = 71: 24: 4: 1. That is,
Sulfur (S) derived from a sulfonic acid group was detected on the bead surface.

When 1.0 g of the beads was placed in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 4.8 mg of albumin was adsorbed on the beads. did.

Similarly, an adsorption test was performed on γ-globulin, and it was found that 1.3 mg of γ-globulin was adsorbed on the beads.

Similarly, an adsorption test was performed on lysozyme, and it was found that 18.1 mg of lysozyme was adsorbed on the beads.

As a result of the above adsorption test, it was found that the porous beads having a sulfonic acid group exhibited selective adsorption to lysozyme.

Example 4 <Preparation of Polymerizable Solution> As the polymerizable compound (A), a urethane acrylate having an average of three acrylic groups in one molecule (manufactured by Dainippon Ink and Chemicals, Inc.) Unidick S9-414) 95 parts, dimethylaminoethyl methacrylate quaternary compound (Kyoeisha Chemical Co., Ltd., Light Ester DQ) as a polymerizable compound (B) having a functional functional group
-75) 5 parts, Irgacure 1 as a photopolymerization initiator
84 (manufactured by Ciba-Geigy), 2-propanol (manufactured by Wako Pure Chemical Industries, Ltd.) 100 as a phase separating agent (C)
And a mixed solution of 41 parts of distilled water was uniformly mixed to obtain a polymerizable solution (4).

<Preparation of porous beads> Inner diameter 0.16 mm
And the polymerizable solution (4) was 22 ml / min (discharge linear velocity 1833 cm / sec).
In the same manner as in Example 1, except that the beads were extruded, white beads having no gloss on the surface were obtained.

<Characteristics of beads> The beads have a diameter of 0.5
When observed with a scanning electron microscope, both the outer surface and the cut interior had a porous structure in which polymer fine particles adhered to each other. The gap between the particles is about 2μm
Met.

The element ratio on the surface of the beads is C: O: N
= 51:21:28. That is, nitrogen (N) derived from a quaternary ammonium base was detected on the bead surface.

When 1.0 g of the beads was placed in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 1.0 mg of albumin was adsorbed on the beads. did.

When a similar adsorption experiment was performed using a 0.3% bovine serum albumin aqueous solution (pH 3.5), it was found that 0.15 mg of albumin was adsorbed on the beads.

From the results of the above adsorption test, it was found that porous beads having a quaternary ammonium base having a positive charge exhibited selective adsorption to albumin (pH 7.0) having a negative charge. did.

In addition, 1.0 g of the beads was added to 500 EU.
(Endotoxin unit) / 100 ml of an endotoxin aqueous solution was added, and the mixture was stirred at room temperature for 15 minutes. The endotoxin residual amount in the aqueous solution was measured using a wale reader SK601 manufactured by Seikagaku Corporation.
005 EU / dl. That is, 499.9995
It was found that EU endotoxin was adsorbed on the beads.

Further, 1.0 g of the beads was added to 500 mg
/ L bilirubin (causative agent of jaundice) aqueous solution (pH 1)
3) The mixture was placed in 100 ml and stirred at room temperature for 2 hours. As a result, it was found that 21.6 mg of bilirubin was adsorbed on the beads.

Example 5 <Preparation of Polymerizable Solution> As the polymerizable compound (A), caprolactone-modified tris (acryloyloxyethyl) isocyanurate (Aronix M-, manufactured by Toagosei Co., Ltd.)
325) 90 parts, as a polymerizable compound (B) having a functional functional group, 10 parts of EO-modified phthalic acrylate (manufactured by Toagosei Co., Ltd., Aronix M-5400), and as a photopolymerization initiator, Irgacure 184 ( A mixed solution of 2 parts of Ciba-Geigy Co., Ltd., 100 parts of 2-propanol (manufactured by Wako Pure Chemical Industries, Ltd.) and 41 parts of distilled water as a phase separating agent (C) is uniformly mixed, and a polymerizable solution (5 ) Got.

<Production of Porous Beads> In the same manner as in Example 1, white beads having no gloss on the surface were obtained.

<Characteristics of beads> The beads had a diameter of 0.9.
When observed with a scanning electron microscope, both the outer surface and the cut interior had a porous structure in which polymer fine particles adhered to each other. The gap between the particles is about 2μm
Met.

When 1.0 g of the beads was placed in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 5.25 mg of albumin was adsorbed on the beads. did.

From the above results, it was found that the porous beads having a carboxyl group exhibited weak adsorption to proteins and could be used for immunosensors and the like on which an antigen (a type of protein) was immobilized.

Example 6 <Preparation of Polymerizable Solution> As the polymerizable compound (A), a urethane acrylate having a weight average molecular weight of 2,000 and an average of three acryl groups per molecule (Dainippon Ink Chemical Industry) 64 parts of Unidick V-4263), dicyclopentenyl diacrylate (manufactured by Nippon Kayaku Co., Ltd.)
Cayarad R-684) 16 parts, 6- (meth) acryloyl (1-O-) n-butyl glucoside as a polymerizable compound (B) having a functional functional group, 20 parts, Irgacure 184 as a photopolymerization initiator (Manufactured by Ciba Geigy)
2 parts, as a phase separating agent (C), 100 parts of polyvinylpyrrolidone having a molecular weight of 10,000 (Tokyo Kasei Kogyo Co., Ltd.) and N, N-dimethylacetamide (Wako Pure Chemical Industries, Ltd.)
(Product) (300 parts) was uniformly mixed to obtain a polymerizable solution (6).

<Production of Porous Beads> In the same manner as in Example 1, white beads having no gloss on the surface were obtained.

<Characteristics of beads> The beads had a diameter of 0.9.
When observed with a scanning electron microscope, it was found that both the outer surface and the cut interior had a polymer network porous structure. The pore size of the mesh on the surface was about 1 μm, and the pore size of the cut inner mesh was about 2 μm.

[0077] 1.0 g of the beads was placed in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours. As a result, it was found that 0.01 mg of albumin was adsorbed on the beads. .

That is, it has been found that porous beads having a glucoside group hardly adsorb protein, so that clogging of beads due to protein adsorption hardly occurs, and it can be used as a biological column.

Example 7 <Preparation of Polymerizable Solution> As the polymerizable compound (A), an epoxy acrylate having a weight average molecular weight of 600 and two acryl groups per molecule (Epoxy acrylate manufactured by Kyoeisha Chemical Co., Ltd. Ester 3002A) 60 parts, ethylene oxide-modified bisphenol A diacrylate (Daiichi Kogyo Seiyaku Co., Ltd., New Frontier BPE-4) 20 parts, Nα- as a polymerizable compound (B) having a functional functional group
20 parts of (methacryloyloxyethylaminocarbonyl) tryptophan, 2 parts of Irgacure 184 (manufactured by Ciba Geigy) as a photopolymerization initiator, and polyvinylpyrrolidone having a molecular weight of 10,000 as a phase separating agent (C) (manufactured by Tokyo Chemical Industry Co., Ltd.) A mixed solution of 100 parts and 320 parts of N, N-dimethylacetamide (manufactured by Wako Pure Chemical Industries, Ltd.) was uniformly mixed to obtain a polymerizable solution (7).

<Preparation of Porous Beads> In the same manner as in Example 1, white beads having no gloss on the surface were obtained.

<Characteristics of Beads> These beads have a diameter of 0.9.
When observed with a scanning electron microscope, both the outer surface and the cut interior had a polymer network porous structure. The pore size of the mesh was about 2 μm.

The element ratio on the surface of the beads is C: O:
N: 74: 21: 5. That is, nitrogen (N) derived from tryptophan residues was detected on the bead surface.

When 1.0 g of the beads was placed in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 5.7 mg of albumin was adsorbed on the beads. did.

Similarly, an adsorption test was carried out on bovine serum γ-globulin. As a result, it was found that 31 mg of γ-globulin was adsorbed on the beads.

As a result of the above adsorption test, it was found that the porous beads having tryptophan residues exhibited selective adsorption to γ-globulin.

Comparative Example 1 <Preparation of Polymerizable Solution> As the polymerizable compound (A), a urethane acrylate having a weight average molecular weight of 2,000 and an average of three acrylic groups per molecule (Dainippon Ink Chemical Industry) 80 parts of Unidick V-4263 manufactured by K.K., hexanediol diacrylate (Daiichi Kogyo Seiyaku Co., Ltd.)
20 parts of New Frontier HDDA), Irgacure 184 (manufactured by Ciba Geigy) as a photopolymerization initiator
2 parts, as phase separator (C), methyl caprate 180
Parts (manufactured by Wako Pure Chemical Industries, Ltd.) were uniformly mixed to obtain a polymerizable solution.

<Production of Porous Beads> In the same manner as in Example 1, white beads having no gloss on the surface were obtained.

<Characteristics of beads> The beads had a diameter of 0.9.
When observed with a scanning electron microscope, the outer surface had a mesh-like structure with a pore diameter of about 1 μm, and the cut inside had a porous structure in which polymer fine particles were adhered to each other. The gap between the fine particles was about 0.8 μm.

When 1.0 g of the beads was placed in 100 ml of a 0.3% bovine serum albumin aqueous solution (pH 7.0) and stirred at room temperature for 2 hours, it was found that 18.1 mg of albumin was adsorbed on the beads. did.

A similar adsorption experiment was performed using a 0.3% bovine serum γ-globulin aqueous solution (pH 7.0), and it was found that 18.8 mg of γ-globulin was adsorbed on the beads. did.

From the above results, it was found that the porous beads having no functional functional group were adsorbed only by the hydrophobic interaction, and thus had no selective adsorption to proteins.

In addition, 1.0 g of the beads was added to 500 EU.
When the mixture was added to 100 ml of an endotoxin aqueous solution and stirred at room temperature for 15 minutes, the endotoxin concentration in the aqueous solution was 483 EU / dl, and almost no adsorbability of the beads to endotoxin was observed.

Furthermore, 1.0 g of the beads was added to 500 mg
/ L bilirubin (causative agent of jaundice) aqueous solution (pH 1)
3) When the mixture was put in 100 ml and stirred at room temperature for 2 hours, the amount of adsorbed bilirubin was 0.1 mg, and almost no adsorbability of beads to bilirubin was observed.

[0094]

According to the production method of the present invention, the type and amount of functional functional groups can be widely adjusted, and several types of functional functional groups can be simultaneously introduced and the particle size can be controlled. Certain porous beads can be efficiently produced.

Claims (5)

[Claims] 1. A crosslinked polymer forming material (A) comprising a polymerizable compound having two or more unsaturated groups polymerizable in one molecule by irradiation with active energy rays, an ionic group, a carboxyl group, a phosphoric acid residue, A polymerizable compound (B) having a functional functional group selected from the group consisting of an amino group, a sugar residue and an amino acid residue and an unsaturated group polymerizable by irradiation with active energy rays; A is formed by dissolving or dissolving in A) or dissolving in a homogeneous solution of the crosslinked polymer-forming material (A) and a solvent and irradiating the crosslinked polymer-forming material (A) with an active energy ray. A uniform polymerizable solution mixed with a phase separator (C) which is incompatible with the cross-linked polymer and which is inactive with respect to active energy rays is discharged in the form of droplets, and the active energy is Irradiate the line and then And removing the phase-separating agent (C) and the unreacted polymerizable solution. 2. The production method according to claim 1, wherein the method of discharging the liquid in the form of droplets is a method in which the polymerizable solution is extruded from a nozzle under the condition that the polymerizable solution becomes droplets during free fall. 3. The method according to claim 1, wherein 1 to 5 parts by weight of the phase separator (C) is used per 1 part by weight of the crosslinked polymer forming material (A). 4. The polymerizable compound (B) is a (meth) acrylate having an ionic group, a (meth) acrylate having a carboxyl group, a phosphoric acid (meth) acrylate, a (meth) acrylate having an amino group, and a sugar (meth). 4. The method according to claim 1, wherein the (meth) acrylic polymerizable compound is selected from the group consisting of acrylates and (meth) acrylates of amino acids. 5. The method according to claim 1, wherein the polymerizable compound (B) is used in an amount of 0.03 to 3 parts by weight based on 1 part by weight of the crosslinked polymer forming material (A).