JP6596766B2 - Sugar-modified polymer material and method for producing the same - Google Patents

Description

  In the present invention, a sugar compound is fixed to a base material made of a polymer material, and functions such as hydrophilicity, wettability, and adsorptivity are imparted. For example, sugars used for cell separation base materials, cell culture base materials, etc. The present invention relates to a modified polymer material and a method for producing the same.

  In recent years, it has become clear that sugar chains are deeply involved in various life phenomena (cell recognition and information transmission, cell adhesion, differentiation / proliferation, canceration, virus infection, blood coagulation, immune reaction, etc.). Research into the development of incorporated functional materials and pharmaceuticals is active.

  In order to develop a sugar chain-containing functional material that expresses a target physiological function, a technique for introducing a sugar chain molecule into a polymer is required. Under such circumstances, various sugar chain-containing polymers have been reported.

For example, as a polymer incorporating a sugar chain such as a monosaccharide, disaccharide, or oligosaccharide,
1. Polyvinyl alcohol type (Patent Document 1)
2. Polyacrylate type (Patent Document 2)
3. Polystyrene mold (Patent Document 3, Patent Document 4, Patent Document 5, Patent Document 6, Patent Document 7)
4. Polyether type (Patent Document 8)
5. Polyethyleneimine type (Patent Document 9, Patent Document 10)
6. Polyamino acid type (Patent Document 11, Patent Document 12, Patent Document 13, Patent Document 14, Patent Document 15)
7. A polyurethane type (Patent Document 16) is known.

  In addition, as a method of directly introducing sugar chain molecules into a general-purpose polymer support, there is a method of modifying the surface of the polymer support by graft polymerization, for example, using graft polymerization using an electron beam irradiation method. A polymer material in which a saccharide is fixed to a polymer substrate is known (see Patent Documents 17 to 20).

JP 63-238105 A JP-A-63-68603 JP-A-7-304788 JP-A-8-253495 JP-A-8-319317 JP 2002-88094 A JP-A-2005-112987 Japanese Patent Laid-Open No. 5-140294 Japanese Patent Laid-Open No. 5-140213 JP 2002-302511 A Japanese Patent Laid-Open No. 5-178986 JP-A-8-337666 JP-A-9-227600 Japanese Patent Laid-Open No. 11-60603 JP 2003-73397 A Japanese Patent Laid-Open No. 11-71391 JP 2011-224350 A JP 2011-245267 A JP 2008-255348 A JP-A-5-310860

  When various sugar chain-containing polymers disclosed in Patent Documents 1 to 16 are used as a cell separation substrate and a cell culture substrate, a film or the like is formed from the sugar chain-containing polymer, or a sugar chain-containing polymer is used. It is necessary to coat the polymer with a polymer support. However, when forming a film or the like, it is necessary to make a sugar chain-containing polymer once, so there is a problem that the production cost becomes high. When the polymer support is coated, the sugar chain-containing polymer is supported. May elute from the body. In order to solve these problems, a technique for directly introducing a sugar chain molecule into a general-purpose polymer support is required. Therefore, Patent Documents 18 to 21 disclose a method for directly introducing a sugar chain molecule into a general-purpose polymer support. In these documents, a polymer base material in which active species (polymer radicals) are generated on the surface by electron beam irradiation, polymerization having an ethylenically unsaturated bond and a sugar chain capable of reacting with the active species (polymer radical). The reactive compound (polymerizable compound having a sugar chain) is brought into contact and reacted in one step, but the reaction yield is very low. That is, the reaction between the polymer substrate and the polymerizable compound is a solid phase-liquid phase reaction between the polymer substrate that is a solid phase and the polymerizable compound solution that is a liquid phase. Since the polymerizable compound has a large molecular weight, the graft polymerization reaction is hardly promoted.

  In view of the above problems with the surface-modified polymer material, the present invention provides a polymer material having a sugar compound immobilized thereon, which is useful as a cell separation substrate or a cell culture substrate. It should be provided in a good and simple way.

  As a result of intensive studies to solve the above-mentioned problems, the inventors of the present invention grafted a radical polymerizable compound onto a base material made of a polymer material by an electron beam irradiation method to obtain a grafted base material, It has been found that the above-mentioned problems can be solved by chemically bonding a sugar compound to the graft chain of the grafting substrate, and the present invention has been completed.

That is, the present invention is as follows.
(1) A step of obtaining a grafted substrate by graft polymerization of a radically polymerizable compound by a radiation graft polymerization method of irradiating an electron beam onto a petri dish-like, sheet-like or fibrous substrate made of a polymer material; A step of immersing the grafted substrate in a sugar compound reaction solution in which the sugar compound is dissolved in a solvent to impart the sugar compound to the grafted substrate, and then chemically bonding the sugar compound to the graft chain of the grafted substrate; A method for producing a sugar-modified polymer material containing
(2) The sugar-modified polymer material according to (1), wherein the sugar-modified polymer material according to (1) is immersed in the sugar compound reaction solution to give a sugar compound to the grafted substrate, and then laminated and heated on both surfaces of the substrate. Production method.
(3) The method for producing a sugar-modified polymer material according to (1) or (2), wherein the solvent contains a tertiary amine.
(4) The method for producing a sugar-modified polymer material according to any one of (1) to (3), wherein the solvent contains an aprotic polar solvent.
(5) The ratio of sugar compound to tertiary amine mol / L [sugar compound (mol / L) / tertiary amine (mol / L)] in the solvent is 0.1 or more and 1.5 or less. The method for producing a sugar-modified polymer material according to (3) or (4).
(6) The method for producing a sugar-modified polymer material according to any one of (1) to (5), wherein the sugar compound is contained in the solvent in an amount of 0.010 mol / L to 5.000 mol / L.
(7) The method for producing a sugar-modified polymer material according to any one of (3) to (6), wherein the solvent contains a tertiary amine in an amount of 0.001 mol / L to 7.500 mol / L.
(8) The method for producing a sugar-modified polymer material according to any one of (3) to (7), wherein the tertiary amine is N-methylmorpholine.
(9) The aprotic polar solvent is at least one selected from the group consisting of acetonitrile, acetone, diethyl ether, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide and hexamethylphosphoric triamide (4) to (8) A method for producing a sugar-modified polymer material according to any one of the above.
(10) The method for producing a sugar-modified polymer material according to any one of (1) to (9), which is obtained by graft polymerization using at least two kinds of the radical polymerizable compounds.
(11) The method for producing a sugar-modified polymer material according to any one of (1) to (10), wherein the radical polymerizable compound is an acrylic monomer.
(12) The method for producing a sugar-modified polymer material according to (11), wherein the acrylic monomer is at least one selected from the group consisting of acrylic acid, methacrylic acid, and glycidyl methacrylate.
(13) The method for producing a sugar-modified polymer material according to any one of (1) to (12), wherein the sugar compound is chemically bonded using at least two types of the sugar compounds.
(14) The method for producing a sugar-modified polymer material according to any one of (1) to (13), wherein the sugar compound is at least one selected from the group consisting of monosaccharides, disaccharides, oligosaccharides, and polysaccharides. .
(15) The method for producing a sugar-modified polymer material according to (14), wherein the sugar compound is an amino sugar.
(16) The method for producing a sugar-modified polymer material according to (15), wherein the amino sugar is at least one selected from the group consisting of glucosamine, mannosamine and galactosamine.
(17) The sugar modified polymer according to any one of (1) to (16), wherein the polymer material is at least one selected from the group consisting of polyolefin, polystyrene, polyester, polyamide, rayon, polyvinyl alcohol, and cellulose. Manufacturing method of molecular materials.
(18) Production of a sugar-modified cell separation filter comprising a step of filling a sugar-modified polymer material obtained by the production method of any one of (1) to (17) into a container having an inlet and an outlet. Method.
(19) A sugar-modified polymer material in which a sugar compound is chemically bonded to a graft chain of a grafted substrate obtained by graft-polymerizing a radically polymerizable compound on a petri dish-like, sheet-like or fibrous substrate made of a polymer material.
(20) The sugar-modified polymer material according to (19), wherein the graft chain is composed of only a radical polymerizable compound.
(21) The sugar-modified polymer material according to (19) or (20), wherein the graft chain is made of a copolymer of at least two kinds of radical polymerizable compounds. (22) The radical polymerizable compound is an acrylic monomer. The sugar-modified polymer material according to any one of (19) to (21).
(23) The sugar-modified polymer material according to (22), wherein the acrylic monomer is at least one selected from the group consisting of acrylic acid, methacrylic acid, and glycidyl methacrylate.
(24) The sugar-modified polymer material according to any one of (19) to (23), wherein at least two kinds of sugar compounds are chemically bonded to the graft chain.
(25) The sugar-modified polymer material according to any one of (19) to (24), wherein the sugar compound is at least one selected from the group consisting of monosaccharides, disaccharides, oligosaccharides, and polysaccharides.
(26) The sugar-modified polymer material according to (25), wherein the sugar compound is an amino sugar.
(27) The sugar-modified polymer material according to (26), wherein the amino sugar is at least one selected from the group consisting of glucosamine, mannosamine and galactosamine.
(28) The sugar modified polymer according to any one of (19) to (27), wherein the polymer material is at least one selected from the group consisting of polyolefin, polystyrene, polyester, polyamide, rayon, polyvinyl alcohol, and cellulose. Molecular material.
(29) A cell separation material comprising the sugar-modified polymer material according to any one of (19) to (28).
(30) A cell separation filter obtained by filling the sugar-modified polymer material according to any one of (19) to (28) into a container having an inlet and an outlet.

  According to the present invention, a sugar compound is added to a grafted substrate obtained by graft polymerization of a radical polymerizable compound by electron beam irradiation to a substrate made of a polymer material, thereby allowing the sugar compound to react. The fixed polymer material can be provided by a simple method with good reaction yield. The sugar-modified polymer material of the present invention is suitable for cell separation materials, cell culture materials, and other applications due to various functions such as hydrophilicity, wettability, and adsorptivity imparted according to the type of bound sugar compound. Can be used.

  Hereinafter, embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described below.

  The method for producing a sugar-modified polymer material according to the present invention includes graft-polymerizing a radical polymerizable compound by a radiation graft polymerization method of irradiating an electron beam onto a petri dish-like, sheet-like or fibrous base material made of a polymer material. A step of obtaining a grafted substrate, and immersing the grafted substrate in a sugar compound reaction solution in which a sugar compound is dissolved in a solvent to impart the sugar compound to the grafted substrate, and then grafting the grafted substrate. Chemically bonding a sugar compound to the chain. As a result, the sugar-modified compound of the present invention, in which a sugar compound is chemically bonded to a graft chain of a grafted substrate obtained by graft-polymerizing a radically polymerizable compound to a petri dish-like, sheet-like or fibrous substrate made of a polymer material. A molecular material is obtained.

<Step of obtaining grafted substrate>
In the present invention, a grafted substrate is obtained by first graft-polymerizing a radically polymerizable compound onto a petri dish-like, sheet-like or fibrous substrate made of a polymer material by a radiation graft polymerization method in which an electron beam is irradiated.

  In the radiation graft polymerization method of irradiating the electron beam, after generating an active species by irradiating the substrate with an electron beam in advance, a radical polymerizable compound is applied to the substrate on which the generated species is generated, and then After so-called pre-irradiation method that promotes the polymerization of the graft-polymerizable compound by post-polymerization, and after giving a radical-polymerizable compound to the base material, this is irradiated with an electron beam to generate active species, and then post-polymerization There is a so-called simultaneous irradiation method in which the polymerization of the graft polymerization compound is accelerated. In the present invention, both the pre-irradiation method and the simultaneous irradiation method can be employed.

  For the production of the grafted substrate by the pre-irradiation method, for example, a method described in JP-A-2005-60555 can be employed. In addition, for the production of the grafted substrate by the simultaneous irradiation method, for example, a method described in JP-A No. 2001-164467 can be employed.

  In any of the pre-irradiation method and the simultaneous irradiation method, a film may be laminated on the surface of the base material until the post-polymerization is completed after the radical polymerizable compound is applied to the base material. preferable. As a result, volatilization of the radical polymerizable compound can be prevented, graft polymerization is started uniformly, and radical deactivation due to oxygen in the air is suppressed by blocking air. In the case of the simultaneous irradiation method, the surface of the base material is sealed with a film even during electron beam irradiation, and the base material and the radical polymerizable compound are shielded from oxygen in the air. Oxidation of the material is less likely to occur.

  As said film, it is a polymer film which has a thickness of 0.01-0.2 mm, Comprising: The thing of appropriate thickness should just be used according to the permeation | transmission power of the electron beam to be used. Examples of the material include polyesters such as polyethylene terephthalate and polyolefins, but polyethylene terephthalate is preferable. In particular, in the case of the simultaneous irradiation method, a polyethylene terephthalate film having low generation efficiency of polymer radicals by electron beam irradiation and low oxygen permeability is preferable.

  In the pre-irradiation method, first, an active species (polymer radical) that induces radical polymerization is generated by irradiating a base material made of a polymer material with an electron beam. The lower the ambient temperature during electron beam irradiation, the higher the polymer radical generation efficiency, but it may be normal room temperature.

  The electron beam irradiation conditions for the pre-irradiation method are preferably an acceleration voltage of 100 to 2000 kilovolts (hereinafter abbreviated as “kV”), preferably 120 to 120, with the oxygen concentration of the irradiation atmosphere set to 300 ppm or less. In the range of 300 kV and current of 1 to 100 mA, it is determined appropriately according to the thickness of the base material, the type of polymer material, the target graft rate, and the like.

  Further, the irradiation dose of the electron beam may be appropriately determined in consideration of the target graft ratio and a decrease in physical properties of the substrate due to irradiation, and is usually about 10 to 300 kiloGray (hereinafter abbreviated as “kGy”). , Preferably 50 to 200 kGy. When the irradiation dose is less than 10 kGy, the generation of active species necessary for a sufficient amount of graft polymerization does not occur. When the irradiation dose exceeds 300 kGy, the physical properties are reduced due to cleavage of the main chain even when the substrate is made of a radiation-resistant polymer material. It is not preferable because it occurs.

  Note that the irradiation atmosphere in the pre-irradiation method is preferably an inert gas atmosphere such as nitrogen gas, but may be an air atmosphere. However, in an air atmosphere, the base material may be oxidized by oxygen in the air.

  Next, a radical polymerizable compound is imparted to the substrate after the electron beam irradiation. For example, it is immersed in a tank of a radical polymerizable compound solution from which dissolved oxygen has been removed by ventilating nitrogen gas, or dipped through the tank of the radical polymerizable compound solution so as to stay for a predetermined time, and is then applied to the substrate. A radically polymerizable compound is sufficiently imparted.

  In the present invention, “immersion” means that the base material comes into contact with the radical polymerizable compound solution. Therefore, various coating methods can be used as a method for imparting the radical polymerizable compound to the substrate. Among them, an impregnation coat, a comma direct coat, a comma reverse coat, a kiss coat, a gravure coat and the like are preferable because they can be efficiently coated.

  The base material provided with the radical polymerizable compound is taken out from the solution tank. At that time, it is preferable to laminate a film on the surface of the substrate. For example, when a base material is a sheet form or a fiber form, the base material which provided the radically polymerizable compound solution is pinched | interposed between two films, and it adheres. By sticking the film to the substrate to which the radical polymerizable compound solution is applied, the application rate of the radical polymerizable compound solution can be controlled to be constant, and the radical polymerizable compound can be uniformly applied to the substrate. Moreover, the graft reaction is further promoted by reacting the base material and the radical polymerizable compound in a space sealed by laminating the films. The “laminate” in the present invention means that the substrate and the film are brought into contact with each other.

  The base material taken out from the solution tank is allowed to stay in the post-polymerization tank for a predetermined time at a predetermined temperature, thereby promoting graft polymerization of the radical polymerizable compound (post-polymerization). The post-polymerization temperature at this time is 0 degreeC-130 degreeC, More preferably, it is 40 degreeC-70 degreeC. Thereby, the graft polymerization reaction between the substrate and the radical polymerizable compound is promoted. Thereafter, the grafted substrate can be obtained by washing and drying. The post-polymerization atmosphere is preferably an inert gas atmosphere such as nitrogen gas. However, when post-polymerization is performed with the film laminated, an air atmosphere may be used.

  Further, in the case of the simultaneous irradiation method, it is immersed in a tank of a radical polymerizable compound solution from which dissolved oxygen has been removed by ventilating nitrogen gas, or is retained for a predetermined time by passing through the tank, A radically polymerizable compound is sufficiently imparted to the material. Then, it removes from the said solution tank and irradiates with an electron beam. When taking out from a solution tank, it is preferable to laminate | stack a film on the surface of a base material and to irradiate an electron beam in this state.

  The acceleration voltage in the case of the simultaneous irradiation method may be appropriately determined according to the type of polymer material, the total thickness of the substrate and the laminated film provided with the radical polymerizable compound solution, and the target graft ratio, An acceleration voltage of about 100 to 2000 kV is appropriate. Further, the irradiation dose of the electron beam may be the same as that in the pretreatment method. The atmosphere during electron beam irradiation is preferably an inert gas atmosphere such as nitrogen or helium. However, when a film is laminated on the surface of the substrate, there is no effect on the graft polymerization by the irradiation atmosphere. Therefore, irradiation in air is appropriate.

  The substrate irradiated with the electron beam is subjected to post-polymerization in the same manner as in the pre-irradiation method. Thereafter, the grafted substrate can be obtained by washing and drying. In the simultaneous irradiation method, the post-polymerization atmosphere is preferably an inert gas atmosphere such as nitrogen gas. However, when post-polymerization is performed with the film laminated, an air atmosphere may be used.

  The polymer material used in the present invention is not particularly limited as long as it generates an active species (polymer radical) for radical reaction by electron beam irradiation. Examples of the polymer material include polyolefins such as polyethylene or polypropylene, polyesters such as polyethylene terephthalate, polyamides such as nylon, rayon, polyvinyl alcohol, and cellulose.

  Further, when the sugar-modified polymer material of the present invention is used for a leukocyte removal filter material, the polymer material as a base material is not particularly limited as long as it does not easily damage blood cells, and as a specific example, Polyester, polyolefin, polyacrylonitrile, polyamide, polystyrene, polyalkyl methacrylate, polyvinyl chloride, polychloroprene, polyurethane, polyvinyl alcohol, polyvinyl acetate, polysulfone, polyethersulfone, polybutadiene, butadiene-acrylonitrile copolymer, styrene-butadiene copolymer Examples of the polymer include ethylene, vinyl alcohol copolymer, cellulose diacetate, and ethyl cellulose.

  Although the form of the base material made of the polymer material is not particularly limited, examples thereof include a petri dish shape, a sheet shape, and a fiber shape. When the substrate is fibrous, any of a woven fabric, a non-woven fabric and the like may be used. When the sugar-modified polymer material of the present invention is used for a cell separation material such as a filter material, a woven fabric or a non-woven fabric is preferable, and a non-woven fabric is more preferable. Here, the non-woven fabric refers to a cloth-like material in which fibers or a collection of fibers are chemically, thermally, or mechanically bonded regardless of knitting. When a certain shape is maintained, for example, by friction caused by contact between the fibers and the fibers, or by being entangled with each other, the fibers are mechanically coupled.

When the substrate is fibrous, the graft ratio improves as the average fiber diameter is smaller. The “average fiber diameter” is obtained by sampling a part of the fiber material, taking a scanning electron micrograph, measuring the diameters of 100 or more randomly selected fibers, and averaging them. Value. In the present invention, the “grafting ratio” is a value calculated as follows from the dry weight of the base material before the graft reaction (W ₁ ) and the dry weight of the grafted base material after the graft reaction (W ₂ ). .
Graft ratio = [(W ₂ −W ₁ ) / W ₁ ] × 100 (%)

  When the sugar-modified polymer material of the present invention is used as a cell separation material such as a filter material, the average fiber diameter is preferably 1.0 to 20 μm. If it is less than 1.0 μm, it is difficult to produce a nonwoven fabric stably, and the viscosity resistance of the cell-containing liquid tends to be too high. On the other hand, when it exceeds 20 μm, the cell removal ability tends to be low.

  Moreover, the radically polymerizable compound used in the present invention is a compound that forms a bond with a polymer radical (active species) generated on a substrate by electron beam irradiation. Specifically, unsaturated compounds having acidic groups such as acrylic acid, methacrylic acid, itaconic acid, methacryl sulfonic acid, styrene sulfonic acid and the like, esters thereof, unsaturated carboxylic amides such as acrylamide and methacrylamide, glycidyl at the terminal Examples thereof include unsaturated compounds having a group and a hydroxyl group, unsaturated organic phosphates such as vinyl phosphonate, methacrylic acid esters having basicity such as quaternary ammonium salts and tertiary ammonium salts, fluoroacrylates, and acrylonitrile. However, it is not limited to these. These can be used alone or in admixture of two or more. By using two or more kinds of radically polymerizable compounds, a composite grafted substrate comprising a copolymer of at least two kinds of radically polymerizable compounds having a graft chain can be obtained.

  Among these radical polymerizable compounds, in the present invention, it is preferable to use an acrylic monomer from the viewpoint of the graft ratio. Furthermore, from the viewpoint of reactivity with the sugar compound, an acrylic monomer having a carboxy group or an epoxy group at the molecular end is preferable, and more preferably at least one selected from the group consisting of acrylic acid, methacrylic acid, and glycidyl methacrylate. It is.

  An acrylic monomer having a carboxy group at the molecular end, such as acrylic acid or methacrylic acid, or an acrylic monomer having an epoxy group at the molecular end, such as glycidyl methacrylate, is an ethylenic monomer at one end of the monomer molecule. The saturated group facilitates graft polymerization to a base material made of a polymer material, and in the coupling reaction with a sugar compound described later, a carboxy group or an epoxy group at the other end of the monomer molecule and the sugar compound The hydroxy group and the amino group of the amino sugar react well, and the reaction yield is excellent.

  The radical polymerizable compound may be a diluted solution using water, an organic solvent such as a lower alcohol, or a mixed solution thereof as a solvent. The concentration of the radically polymerizable compound in the diluted solution varies depending on the desired graft ratio, but can be prepared at 1 to 70% by volume. Moreover, when using the radically polymerizable compound which is easy to produce | generate a homopolymer, you may suppress the production | generation of a homopolymer by adding copper and iron metal salt to the diluted solution of a radically polymerizable compound. The lower limit of the concentration of the radical polymerizable compound in the solution is preferably 1% by weight or more, more preferably 2.5% by weight or more, further preferably 5% by weight or more, and particularly preferably 10% by weight or more. In addition, the upper limit value of the concentration of the radical polymerizable compound is preferably 70% by weight or less, more preferably 60% by weight or less, further preferably 50% by weight or less, and particularly preferably 40% by weight or less.

  Moreover, when the said radically polymerizable compound solution contains a solvent, a graft rate will improve. In this case, the concentration of the emulsifier in the solvent is preferably adjusted to be in the range of 0.1 to 5% by weight. As the emulsifier, for example, polysorbate is preferable, and examples of the polysorbate include polysorbate 20, 60, 65, 80, etc. Among them, polysorbate 20 having high hydrophilicity is more preferable.

  In addition, it is desirable to remove dissolved oxygen in advance by blowing an inert gas such as nitrogen gas into the radical polymerizable compound solution.

<Step of chemically bonding a sugar compound to the graft chain of the grafting substrate>
The grafted substrate obtained as described above is immersed in a sugar compound reaction solution in which a sugar compound is dissolved in a solvent to impart the sugar compound to the grafted substrate, and then the sugar is added to the graft chain of the grafted substrate. The sugar-modified polymer material of the present invention can be obtained by chemically bonding the compounds.

  Thus, in the present invention, a radically polymerizable compound is first graft-polymerized onto a base material made of a polymer material to obtain a grafted base material, and then a sugar compound is fixed to the obtained grafted base material. Compared to the case where a radically polymerizable compound having a sugar chain is directly reacted with a base material made of a polymer material because the reaction is directly performed in a phase (grafted base material) -liquid phase (sugar compound reaction liquid) reaction. The reaction yield of is increased. In addition, the graft chain (main chain) of the sugar-modified polymer material obtained is composed only of radically polymerizable compounds.

In the present invention, the “reaction yield” means the dry weight of the grafted substrate (W ₃ ) before the reaction with the sugar compound reaction solution, the dry weight of the sugar-modified polymer material (W ₄ ) after the reaction, and It is a value calculated as follows from the assumed maximum sugar compound weight (W ₅ ) that reacts with the functional group of the graft chain.
Reaction yield (%) = [(W ₄ −W ₃ ) / W ₅ ] × 100

The “assumed maximum sugar compound weight (W ₅ ) that reacts with the functional group of the graft chain” is calculated from the grafting rate of the grafted base material to react with the radical polymerizable compound. It can be determined by calculating the amount of sugar compound.

Examples of the method for imparting the sugar compound to the grafted substrate include immersing the saccharide compound reaction solution in which the saccharide compound is dissolved in a solvent, or immersing the saccharide compound reaction solution in which the saccharide compound is dissolved in the solvent. The sugar compound is imparted to the grafted substrate by allowing it to pass for a predetermined time. Subsequently, after taking out from the said solution tank, reaction is accelerated | stimulated at predetermined temperature and time, and a sugar compound is chemically bonded to the graft chain of the grafting base material. Thereafter, the sugar-modified polymer material of the present invention can be obtained by washing and drying.
The “dissolution” in the present invention means that at least a part of the solute is dissolved in the solvent. Therefore, a heterogeneous sugar compound solution may be used for the reaction, and it is more preferable to use a uniform solution for the reaction from the viewpoint of quality control.

  When heating and reacting the grafted substrate to which the sugar compound has been added, a film is laminated on the surface of the grafted substrate to which the sugar compound has been added in the same manner as the graft polymerization reaction in the step of obtaining the grafted substrate. It is preferable to do. For example, when the grafted substrate is in the form of a sheet or fiber, the grafted substrate to which the sugar compound reaction solution is applied is sandwiched between two films and adhered. By adhering the film to the grafted substrate to which the sugar compound reaction solution has been applied, for example, when the substrate is fibrous, such as a woven or non-woven fabric, the penetration of the sugar compound reaction solution into the substrate is promoted In addition, the application rate of the sugar compound reaction solution can be controlled to be constant, and the sugar compound reaction solution can be uniformly applied to the substrate. Moreover, reaction is further accelerated | stimulated by laminating a film and making a grafting base material and a sugar compound react in the sealed space.

  The film may be the same as that used in the graft polymerization. For example, a polyethylene terephthalate film having a thickness of 0.01 to 0.2 mm is suitable.

  The sugar compound used in the present invention is not particularly limited, and for example, at least one selected from the group consisting of monosaccharides, disaccharides, oligosaccharides and polysaccharides can be used, depending on the use of the sugar-modified polymer material. What is necessary is just to select suitably the sugar compound which can provide a function. Moreover, amino sugar can also be used as saccharides.

  One type of sugar compound can be used alone, or two or more types can be used in combination. When two or more kinds of different sugar compounds are used, at least two kinds of sugar compounds are chemically bonded to the graft chain, and a complex sugar-modified polymer material to which two or more different functions are imparted by the respective sugar compounds is obtained. Can do.

  The chemical bond between the graft chain of the grafted substrate and the sugar compound is a functional group possessed by the radical polymerization reactive compound constituting the graft chain of the grafted substrate, such as a carboxy group of acrylic acid or methacrylic acid, or glycidyl methacrylate. By reaction of epoxy groups with hydroxy groups of sugar compounds. Thereby, a sugar-modified polymer material in which a sugar compound is fixed to a base material is obtained.

  In addition, when an amino sugar is used as the saccharide, the sugar compound is also grafted by a chemical bond between the amino group in the amino sugar and the functional group of the radical polymerization reactive compound constituting the graft chain of the graft base. It can also be chemically bonded to the graft chain of the substrate. For example, a glycidyl group of glycidyl acrylate and an amino group of an amino sugar are chemically bonded, and the amino sugar is bonded to a graft chain of a grafting substrate made of a polymer material.

  Examples of the amino sugar include glucosamine, mannosamine, galactosamine, N-acetylglucosamine, N-acetylmannosamine, N-acetylgalactosamine and the like. Among these, at least one selected from the group consisting of glucosamine, mannosamine and galactosamine is preferably used, and glucosamine is more preferable in terms of easy availability.

  The lower limit of the concentration of the sugar compound in the solvent of the sugar compound reaction solution is preferably 0.010 mol / L or more, more preferably 0.100 mol / L or more, and further preferably 0.400 mol / L or more. Moreover, as an upper limit of the density | concentration of a sugar compound, 5.000 mol / L or less is preferable, 2.000 mol / L or less is more preferable, 1.000 mol / L or less is more preferable. By containing a saccharide compound in the solvent in an amount within the above range, the saccharide compound and the grafted substrate react with each other with a high yield.

  The solvent for the sugar compound reaction solution is not particularly limited as long as it is a solvent that can dissolve the sugar compound without dissolving the grafted substrate, but the solvent preferably contains an aprotic solvent. An aprotic polar solvent is preferable in terms of enhancing the reactivity between the sugar compound and the grafted substrate from the viewpoint of the solubility of the sugar compound and the wettability with the grafted substrate. Examples of the aprotic polar solvent include at least one selected from the group consisting of acetonitrile, acetone, diethyl ether, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, and hexamethylphosphoric triamide. Of these, dimethyl sulfoxide (DMSO) is preferred from the viewpoint of wettability with the grafted substrate, and the reaction yield of the sugar compound is increased.

  The sugar compound reaction solution preferably contains a tertiary amine. The tertiary amine that is a base can increase the nucleophilicity of the sugar compound. When amino sugar is used as the sugar compound, since the amino sugar is usually stabilized as a hydrochloride or sulfate, the sugar compound reaction solution in which the amino sugar is dissolved in a solvent tends to be acidic, but the third is a base. The reaction system is neutralized by the tertiary amine and the reaction is promoted.

  Examples of the tertiary amine include triethylamine, N, N-diisopropylethylamine, N-methylmorpholine, quinuclidine, 1,4-diazabicyclo [2.2.2] octane, pyridine, N, N-dimethyl-4-aminopyridine. , Tetramethylethylenediamine and the like. Among these, N-methylmorpholine is preferable because of its good compatibility with the aprotic polar solvent described above, particularly dimethyl sulfoxide (DMSO).

  The lower limit value of the tertiary amine concentration in the solvent of the sugar compound reaction solution is preferably 0.001 mol / L or more, and more preferably 0.500 mol / L. The upper limit of the tertiary amine concentration is preferably 7.500 mol / L or less, more preferably 1.000 mol / L or less. By containing the tertiary amine in an amount within the above range in the solvent of the sugar compound, the reaction between the sugar compound and the grafted substrate is promoted.

  The lower limit of the mol / L ratio [sugar compound (mol / L) / tertiary amine (mol / L)] between the sugar compound and the tertiary amine in the solvent of the sugar compound solution is 0.1. The above is preferable, 0.2 or more is more preferable, and 0.4 or more is more preferable. The upper limit of the mol / L ratio [sugar compound (mol / L) / tertiary amine (mol / L)] between the sugar compound and the tertiary amine in the solvent is preferably 1.5 or less. 1.0 or less is more preferable, and 0.6 or less is more preferable. When the mol / L ratio between the sugar compound and the tertiary amine is within the above range, the reaction between the sugar compound and the grafted substrate is promoted.

<Sugar modified polymer material>
The sugar-modified polymer material of the present invention can be suitably used as a cell separation material such as a cell separation filter material, a cell culture material such as a cell culture dish.

  In the cell separation filter material comprising the sugar-modified polymer material of the present invention, a sugar compound is fixed to a woven or nonwoven substrate made of the polymer material, and high cell separation performance can be exhibited. In addition, since the sugar compound is fixed to the substrate by chemical bonding, there is no risk of clogging of the filter pores due to coating or the like, and the durability is excellent.

  The cell separation filter of the present invention is obtained by filling the sugar-modified polymer material of the present invention as a filter material into a container having an inlet and an outlet. For example, a sugar-modified polymer material as a filter material is appropriately filled in at least a container having a liquid inlet and outlet. The container may take any form such as a sphere, container, cassette, bag, tube, column, and the like. Preferable specific examples include, for example, a transparent or translucent cylindrical container having a volume of about 0.1 to 1000 ml and a diameter of about 0.1 to 15 cm, or a square or rectangle having a length of about 0.1 to 20 cm. And a rectangular columnar shape having a thickness of about 0.1 to 5 cm. One or more sugar-modified polymer materials as the filter material may be stacked in the flow direction of the cell-containing liquid and filled in the container. Further, another filter material may be further filled on the upstream side and / or the downstream side of the sugar-modified polymer material filled in the container.

  The substrate shape of the sugar-modified polymer material when used as a filter material as described above preferably has a large surface area from the viewpoint of contact frequency in order to make contact with the cell-containing solution in the liquid layer. For example, fibrous structures, such as a woven fabric and a nonwoven fabric, are mentioned. In view of cell adsorbability and handleability as a separating material, woven fabrics and non-woven fabrics are preferred. Nonwoven fabrics are most preferred because they can make multipoint contact with cells.

Example 1
<Manufacture of grafted substrate>
Glycidyl methacrylate was dissolved in a solvent in which water and ethanol were mixed at a ratio of 1: 1 (weight ratio) to prepare a glycidyl methacrylate solution having a monomer concentration of 10% by weight. Dissolved oxygen was removed by bubbling nitrogen gas through the glycidyl methacrylate solution to obtain a radical polymerizable compound solution.
Electron beam irradiation is performed at room temperature in a nitrogen gas atmosphere under conditions of an acceleration voltage of 250 kV and an irradiation dose of 50 kGy on a polypropylene non-woven fabric (average fiber diameter 3 μm, basis weight 40 g / m ² ) of length 14 cm × width 20 cm × thickness 0.43 mm It was.
The nonwoven fabric after electron beam irradiation was immersed in the radical polymerizable compound solution, and a polymerizable compound was imparted to the nonwoven fabric. Polyester films (upper film thickness: 25 μm, lower film thickness: 50 μm) were laminated and adhered to the upper and lower sides of the nonwoven fabric provided with the polymerizable compound. The nonwoven fabric laminated with the film was heated together with the film in an oven at 50 ° C. for 30 minutes to promote the polymerization reaction. Thereafter, the films on both sides of the nonwoven fabric were peeled off, washed with water and methanol, and then dried to obtain a grafted polypropylene nonwoven fabric.
The graft ratio of the obtained grafted polypropylene nonwoven fabric was calculated from the nonwoven fabric dry weight (W ₁ ) before electron beam irradiation and the nonwoven fabric dry weight (W ₂ ) after the graft reaction as follows. It was.
Graft ratio = [(W ₂ −W ₁ ) / W ₁ ] × 100 (%)

<Binding of sugar compound to graft chain of grafted polypropylene nonwoven fabric>
Glucosamine hydrochloride and N-methylmorpholine (NMM) were dissolved in dimethyl sulfoxide (DMSO) to obtain a sugar compound reaction solution having a glucosamine hydrochloride concentration of 0.46 mol / L and an N-methylmorpholine concentration of 0.92 mol / L. .
The grafted polypropylene nonwoven fabric was immersed in the sugar compound reaction solution, and glucosamine hydrochloride and N-methylmorpholine were imparted to the grafted polypropylene nonwoven fabric. A polyester film (upper film thickness: 25 μm, lower film thickness: 50 μm) was laminated on and under the nonwoven fabric and adhered, and the whole film was heated in an oven at 100 ° C. for 10 minutes to promote the reaction ( Hereinafter, this is referred to as a film laminating method.) Thereafter, the films on both sides of the nonwoven fabric were peeled off, washed with water and methanol, and then dried to obtain a sugar-modified polypropylene nonwoven fabric in which glucosamine was chemically bonded to a graft chain composed of glycidyl methacrylate.
In the ATR-IR spectrum of the obtained sugar-modified propylene nonwoven fabric, it was confirmed that a glucosamine-derived OH group peak newly appeared in the vicinity of 3300 cm ⁻¹ .

About the obtained sugar-modified polypropylene nonwoven fabric, the nonwoven fabric dry weight (W ₃ ) before the reaction with the sugar compound reaction solution, the nonwoven fabric dry weight (W ₄ ) after the reaction, and the assumed maximum glucosamine amount that reacts with the epoxy group of the graft chain ( From W ₅ ), the reaction yield of glucosamine was calculated as follows and found to be 2%.
Reaction yield (%) = [(W ₄ −W ₃ ) / W ₅ ] × 100

(Example 2)
A sugar-modified polypropylene nonwoven fabric was obtained by a film laminating method in the same manner as in Example 1 except that the reaction time with the sugar compound solution was 60 minutes. The reaction yield of glucosamine was 19%.

(Example 3)
A sugar-modified polypropylene nonwoven fabric was obtained by a film laminating method in the same manner as in Example 1 except that the reaction time with the sugar compound solution was 300 minutes. The reaction yield of glucosamine was 29%.

Example 4
The grafted polypropylene nonwoven fabric obtained in the same manner as in Example 1 was immersed in the same sugar compound reaction solution as in Example 1, and the reaction solution was heated at 100 ° C. for 300 minutes to promote the reaction (solution method). Thereafter, the nonwoven fabric was taken out from the reaction solution, washed with water and methanol, and then dried to obtain a sugar-modified polypropylene nonwoven fabric in which glucosamine was bonded to a graft chain composed of glycidyl methacrylate. The reaction yield of glucosamine in the obtained sugar-modified polypropylene nonwoven fabric was 10%.

  Table 1 shows the production conditions and results of the sugar-modified polymer materials of Examples 1 to 4 described above.

Claims (25)

  A step of obtaining a grafted substrate by graft-polymerizing a radically polymerizable compound onto a petri dish-like, sheet-like or fibrous substrate made of a polymer material by a radiation graft polymerization method irradiating an electron beam; and the grafted group The material is immersed in a sugar compound reaction solution in which a sugar compound is dissolved in a solvent to give the sugar compound to the grafted substrate, and then a film is laminated on both surfaces of the substrate, heated, And a step of chemically bonding a sugar compound to the graft chain.   The method for producing a sugar-modified polymer material according to claim 1, wherein the solvent contains a tertiary amine.   The method for producing a sugar-modified polymer material according to claim 1 or 2, wherein the solvent contains an aprotic polar solvent. The ratio [sugar compound (mol / L) / tertiary amine (mol / L)] between the sugar compound and the tertiary amine in the solvent is 0.1 or more and 1.5 or less. Item 4. The method for producing a sugar-modified polymer material according to Item 2 or 3.   The method for producing a sugar-modified polymer material according to any one of claims 1 to 4, wherein a sugar compound is contained in the solvent in an amount of 0.010 mol / L or more and 5.000 mol / L or less.     The method for producing a sugar-modified polymer material according to any one of claims 2 to 5, wherein the solvent contains a tertiary amine in an amount of 0.001 mol / L to 7.500 mol / L.   The method for producing a sugar-modified polymer material according to any one of claims 2 to 6, wherein the tertiary amine is N-methylmorpholine.   The aprotic polar solvent is at least one selected from the group consisting of acetonitrile, acetone, diethyl ether, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, and hexamethylphosphoric triamide. A method for producing the sugar-modified polymer material described in 1.   The method for producing a sugar-modified polymer material according to any one of claims 1 to 8, which is obtained by graft polymerization using at least two kinds of the radical polymerizable compounds.   The method for producing a sugar-modified polymer material according to any one of claims 1 to 9, wherein the radical polymerizable compound is an acrylic monomer.   The method for producing a sugar-modified polymer material according to claim 10, wherein the acrylic monomer is at least one selected from the group consisting of acrylic acid, methacrylic acid, and glycidyl methacrylate.   The method for producing a sugar-modified polymer material according to any one of claims 1 to 11, wherein the sugar-modified polymer material is chemically bonded using at least two kinds of the sugar compounds.   The method for producing a sugar-modified polymer material according to any one of claims 1 to 12, wherein the sugar compound is at least one selected from the group consisting of monosaccharides, disaccharides, oligosaccharides, and polysaccharides.   The method for producing a sugar-modified polymer material according to claim 13, wherein the sugar compound is an amino sugar.   The method for producing a sugar-modified polymer material according to claim 14, wherein the amino sugar is at least one selected from the group consisting of glucosamine, mannosamine and galactosamine.   The sugar-modified polymer material according to any one of claims 1 to 15, wherein the polymer material is at least one selected from the group consisting of polyolefin, polystyrene, polyester, polyamide, rayon, polyvinyl alcohol, and cellulose. Production method.   A method for producing a cell separation filter comprising a step of filling a sugar-modified polymer material obtained by the production method according to any one of claims 1 to 16 into a container having an inlet and an outlet. A sugar compound is directly chemically bonded to the graft chain of a grafted substrate obtained by graft-polymerizing a radically polymerizable compound on a petri dish, sheet or fiber substrate made of a polymer material, and the graft chain is only a radically polymerizable compound. consists, the radical polymerizable compound is at least one acrylic monomer selected from the group consisting of acrylic acid, methacrylic acid and glycidyl methacrylate, wherein the sugar compound is a monosaccharide, disaccharide and oligosaccharide or al A sugar-modified polymer material that is at least one selected from the group consisting of:
  The sugar-modified polymer material according to claim 18, wherein the graft chain is made of a copolymer of at least two kinds of radically polymerizable compounds.   The sugar-modified polymer material according to claim 18 or 19, wherein at least two kinds of sugar compounds are chemically bonded to the graft chain.   The sugar-modified polymer material according to any one of claims 18 to 20, wherein the sugar compound is an amino sugar.   The sugar-modified polymer material according to claim 21, wherein the amino sugar is at least one selected from the group consisting of glucosamine, mannosamine and galactosamine.   The sugar-modified polymer material according to any one of claims 18 to 22, wherein the polymer material is at least one selected from the group consisting of polyolefin, polystyrene, polyester, polyamide, rayon, polyvinyl alcohol, and cellulose.   A cell separation material comprising the sugar-modified polymer material according to any one of claims 18 to 23. A cell separation filter formed by filling the sugar-modified polymer material according to any one of claims 18 to 23 into a container having an inlet and an outlet.