Chromatography material raw material composition, chromatography material and its preparation method and application, monolithic column and chromatography stack

Abstract

The present invention relates to a chromatography material raw material composition, a chromatography material, its preparation method and applications, a monolithic column, and a chromatography stack. The chromatography material raw material composition contains 20% to 70% of the raw material A, 30% to 80% of the porogen B, and 0.1‰ to 1‰ of the catalyst C, where the total weight of raw material A and porogen B is 100%. Raw material A contains a polyepoxy group-containing substance. Chromatography materials prepared with the chromatography material composition of the present invention can solve the current problems associated with the application of chromatography microsphere fillers to biopolymers. The chromatography material of the present invention has the characteristics of a uniform internal structure, high preparation reproducibility, high toughness, the ability to be modified with functional ligands, and relatively high hydrophilicity, making it particularly suitable for the separation and purification of biomolecules.

Description

[Technical Field]

[0001] This application claims priority from Chinese Patent Application No. 2022116009496, filed December 12, 2022. This application is incorporated herein by reference in its entirety.

[0002] Specifically, the present invention relates to a chromatography material raw material composition, a chromatography material and its preparation method and application, a monolithic column and a chromatography stack. [Background technology]

[0003] Currently, the separation and purification of biomolecules is basically performed using chromatographic microsphere media with surface ligand modification to achieve the separation and purification of different functions, obtaining relatively pure molecules for use in medicines, diagnostic reagents, biochemical raw materials, food, cosmetic additives, etc. Chromatographic microspheres have become a significant bottleneck in the field of biopolymer purification, including plasmid DNA, mRNA, viral vectors, exosomes, antibody-coupled drugs (ADCs), and complex macromolecular proteins. Specifically, the shortcomings are as follows:

[0004] 1. The pore distribution of the chromatography microspheres is small, and some macromolecules cannot effectively enter the middle and deep parts of the pores, resulting in low ligand utilization and low throughput.

[0005] 2. The porous structure of chromatography microspheres has obvious diffusion effect. To fully diffuse on the pore surface of porous microspheres, the retention time needs to be increased, which results in a long process cycle.

[0006] 3. The internal pore structure of the chromatography microsphere structure is prone to eddy current effects, which generate relatively large shear forces. In addition, the porous structure of the chromatography microspheres contains dead holes, which make them prone to local binding and difficult to dissolve. This results in a relatively low yield of active biopolymers (such as lentiviruses, herpes viruses, exosomes, and relatively large nucleic acid molecules), even as low as 10-20%.

[0007] 4. Chromatographic microspheres need to be filled during use to prevent breakage and contamination, which will unwittingly increase the processing costs and batch-to-batch stability risks of biopharmaceuticals.

[0008] 5. The production cost of chromatography microspheres themselves is relatively high, and batch-to-batch stability of microspheres has always been a problem for domestic chromatography fillers. Controlling the batch stability of microspheres will indirectly increase the processing costs of microspheres, ultimately increasing the downstream production costs of biopharmaceutical companies.

[0009] Monolithic columns, as a fourth-generation chromatography column, have attracted widespread attention in recent years as a separation material. However, existing polymer monolithic columns are prone to forming non-uniform internal structures during the manufacturing process, resulting in uneven distribution of surface functional groups, poor adsorption performance, non-uniform structure, and poor reproducibility. These disadvantages result in low separation column efficiency and limit their application in analytical separation.

[0010] Therefore, there is an urgent need to develop new monolithic column materials to overcome the above-mentioned defects. Summary of the Invention [Problem to be solved by the invention]

[0011] The technical problem to be solved by the present invention is to overcome the drawbacks of existing chromatography microspheres in the field of biopolymer purification, such as low throughput, long process cycle, low yield of effective active biopolymers, low lot-to-lot stability, and high manufacturing costs, and to overcome the drawbacks of existing monolithic columns, such as heterogeneity in the internal structure and poor reproducibility, and to provide a chromatography material raw material composition, a chromatography material and its preparation method and application, a monolithic column, and a chromatography stack. [Means for solving the problem]

[0012] The present invention provides the following technical solutions to solve the above technical problems.

[0013] The present invention provides a chromatography material raw material composition, the total weight of raw material A and porogen B being 100%, 20% to 70% of raw material A, 30% to 80% of Porogen B, 0.1‰ to 1‰ of catalyst C; and The raw material A includes a polyepoxy group-containing substance.

[0014] In the present invention, the polyepoxy group-containing substance refers to a substance containing two or more epoxy groups in its structure.

[0015] In the present invention, the polyepoxy group-containing substance is preferably a polyepoxy glycidyl ether-based substance or a polyepoxy glycidyl ester-based substance.

[0016] In the present invention, the glycidyl ether-based material may be one or more of the monomers satisfying the following structural formula I: [ka] In structural formula I, R 1 is a hydrogen atom, a substituted or unsubstituted C1-C 10 or an epoxy group, n is a positive integer from 0 to 10, L 1 is selected from an oxygen atom or a nitrogen atom, L 2 is selected from an oxygen atom or a nitrogen atom, X is a substituted or unsubstituted C1-C 10 Alkylene group, substituted or unsubstituted C3-C 10 a cycloalkylene group of the formula: [ka] or a benzene ring.

[0017] Preferably, the glycidyl ether-based substance is one or more of the following monomers (a) to (m): [ka]

[0018] The above structure (a) is the structure of glycerol triglycidyl ether.

[0019] The above structure (b) is the structure of pentaerythritol tetraglycidyl ether.

[0020] The above structure (j) is the structure of 1,4 butanediol diglycidyl ether.

[0021] The above structure (k) is the structure of trimethylolethane triglycidyl ether.

[0022] The above structure (l) is the structure of bisphenol A diglycidyl ether.

[0023] The above structure (m) is the structure of bisphenol F diglycidyl ether.

[0024] In the present invention, the glycidyl ether-based material may be one or more polymers satisfying the following structural formula II: [ka] In structural formula II, R 2 is a hydrogen atom, a substituted or unsubstituted C1-C 10 or an alkyl group of [ka] is selected from m is a positive integer from 2 to 40.

[0025] Preferably, the glycidyl ether-based substance is one or more of the following polymers (1) to (4): [ka]

[0026] In the present invention, the glycidyl ester-based material may be one or more of the monomers satisfying the following structural formula III: [ka] In structural formula III, Y is a substituted or unsubstituted C1-C 10 Alkylene group, substituted or unsubstituted C3-C 10 or a benzene ring.

[0027] Preferably, the glycidyl ester-based substance is the following monomer (A) and / or (B): [ka]

[0028] In the present invention, the polyepoxy group-containing substance preferably contains one or more of the following monomers (I) to (IV): [ka]

[0029] In the present invention, the polyepoxy group-containing substance is preferably a polyepoxy glycidyl ether monomer and / or a polyepoxy glycidyl ether polymer.

[0030] Here, the polyepoxy glycidyl ether monomer may be glycerol triglycidyl ether and / or pentaerythritol tetraglycidyl ether.

[0031] Here, the polyepoxy glycidyl ether polymer may be polyglycerol glycidyl ether and / or polypentaerythritol tetraglycidyl ether.

[0032] In the present invention, when the polyepoxy group-containing substance is a mixture of the polyepoxy glycidyl ether monomer and the polyepoxy glycidyl ether polymer, the mixing volume ratio of the polyepoxy glycidyl ether monomer to the polyepoxy glycidyl ether polymer may be (0.1 to 9):1, for example, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, or 8:1.

[0033] In the present invention, the molecular weight of the polyepoxy group-containing substance may be 300 to 2000, for example, a molecular weight of 550 or a molecular weight of 700.

[0034] In the present invention, the weight percentage of the raw material A to the total weight of the raw material A and the porogen B is preferably 22% to 70%, for example, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 58%, 65% or 68%.

[0035] In the present invention, the raw material A may further contain a monoepoxy group-containing substance. The monoepoxy group-containing substance refers to a substance containing one epoxy group in its structure.

[0036] The monoepoxy group-containing substance is preferably a monoepoxy glycidyl ether-based substance or a monoepoxy glycidyl ester-based substance.

[0037] Here, the monoepoxy glycidyl ether-based substance may be a substance containing one epoxy group in its structure, which is common in this field, and is preferably phenyl glycidyl ether and / or butyl glycidyl ether.

[0038] Here, the monoepoxy glycidyl ester-based substance may be one or more of phenyl glycidyl ether, butyl glycidyl ether, pentyl glycidyl ether, octyl glycidyl ether, octadecyl glycidyl ether, and naphthyl glycidyl ether, and is preferably phenyl glycidyl ether and / or butyl glycidyl ether.

[0039] In the present invention, the porogen B is one or more chemical solvents compatible with the raw material A and the catalyst C, preferably one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide (DMF), dimethyl sulfoxide (DMSO), and methyl t-butyl ether, more preferably one or more of toluene, dioxane, and methyl t-butyl ether, such as toluene and dioxane, toluene and methyl t-butyl ether, or dioxane and methyl t-butyl ether.

[0040] In the present invention, when the porogen B is a mixture of two different substances, the volume ratio of the two different substances may be (0.1 to 9):1, for example, 0.5:1, 1:1, 2:1, 3:1, 5:1 or 7:1.

[0041] In the present invention, the weight percentage of the porogen B relative to the total weight of the raw material A and the porogen B is preferably 35% to 80%, for example, 40%, 45%, 50%, 60%, 65%, 70%, 75% or 78%.

[0042] In the present invention, the catalyst C may be a Lewis acid and / or a complex of a Lewis acid.

[0043] wherein the Lewis acid may be one or more of aluminum trichloride, boron trifluoride, iron bromide, iron chloride, zinc chloride, niobium trichloride or niobium pentachloride, and sulfur trioxide, such as boron trifluoride.

[0044] wherein the Lewis acid complex may be one or more of boron trifluoride diethyl etherate, boron trifluoride acetonitrile, boron trifluoride dimethyl carbonate, and boron trifluoride ethylamine.

[0045] In the present invention, the weight percentage of the catalyst C relative to the total weight of the raw material A and the porogen B is preferably 0.3‰ to 1‰, for example, 0.4‰, 0.5‰, 0.6‰, 0.7‰, 0.8‰ or 0.9‰.

[0046] In some preferred embodiments of the present invention, the raw material A is a polyepoxy glycidyl ether based monomer and / or a polyepoxy glycidyl ether based polymer, and the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide (DMF), dimethyl sulfoxide (DMSO), and methyl t-butyl ether.

[0047] In some preferred embodiments of the present invention, the raw material A is a polyepoxy glycidyl ether based polymer and a monoepoxy glycidyl ether based monomer, and the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide (DMF), dimethyl sulfoxide (DMSO), and methyl t-butyl ether.

[0048] In some preferred embodiments of the present invention, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide (DMF), dimethyl sulfoxide (DMSO), and methyl t-butyl ether, and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether.

[0049] In some preferred embodiments of the present invention, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, the porogen B is one or more of toluene, dioxane, and methyl t-butyl glyceryl ether, and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether.

[0050] In some preferred embodiments of the present invention, the chromatography material raw material composition is: 50 to 70% of raw material A, which is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, and the monoepoxy glycidyl ether-based monomer is phenyl glycidyl ether and / or butyl glycidyl ether; The composition contains 30 to 50% of porogen B, which is "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane."

[0051] In some preferred embodiments of the present invention, the chromatography material raw material composition is: 50 to 70% of raw material A, which is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, and the monoepoxy glycidyl ether-based monomer is phenyl glycidyl ether and / or butyl glycidyl ether; 30 to 50% of porogen B, which is "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane", 1‰ of catalyst C and the components of

[0052] In some preferred embodiments of the present invention, the chromatography material raw material composition is: 60% of raw material A, which is a polyglycerol triglycidyl ether having a molecular weight of 550 and a butyl glycidyl ether, and the volume ratio of the polyglycerol triglycidyl ether having a molecular weight of 550 to the butyl glycidyl ether is 2:1; 40% Porogen B, wherein the Porogen B is dioxane and / or methyl t-butyl glyceryl ether, and the volume ratio of the dioxane to the methyl t-butyl glyceryl ether is 1:1; 1‰ of catalyst C and the components of

[0053] In one preferred embodiment of the present invention, the total weight of raw material A and porogen B is 100%, and the chromatography material raw material composition is 60% of raw material A, which is a polyglycerol triglycidyl ether having a molecular weight of 550 and a butyl glycidyl ether, and the volume ratio of the polyglycerol triglycidyl ether having a molecular weight of 550 to the butyl glycidyl ether is 2:1; 40% Porogen B, wherein the Porogen B is dioxane and methyl t-butyl glyceryl ether, and the volume ratio of the dioxane to the methyl t-butyl glyceryl ether is 1:1; 1‰ of catalyst C, which is boron trifluoride diethyl etherate; and

[0054] The present invention further provides a chromatography material comprising the aforementioned starting composition, the chromatography material has through-holes distributed therein with an average pore size of 0.2 to 10 μm; The porosity of the through holes is 30 to 80%, the pore volume of the through holes is 0.5 to 2.5 mL / g; The density of the epoxy ligand in the chromatography material is 30 to 80 μmol / mL.

[0055] In the present invention, the average pore size of the through holes is preferably 0.4 to 8 μm, for example, 0.44 μm, 1.4 μm, 0.74 μm, 0.8 μm, 2 μm, 2.7 μm, 3.4 μm, 3.7 μm, 3.8 μm, 4.2 μm, 4.3 μm, 4.5 μm, 5.2 μm, 6 μm, 7 μm, or 7.5 μm.

[0056] In the present invention, the porosity of the through holes is preferably 40 to 68%, for example, 45.9%, 46.5%, 46.8%, 48.8%, 49.7%, 59.7%, 58.6%, 60.1%, 60.4%, 65.1%, 65.9%, or 66.3%.

[0057] In the present invention, the pore volume of the through holes is preferably 0.5 to 1.8 mL / g, for example, 0.53 mL / g, 0.58 mL / g, 0.59 mL / g, 0.62 mL / g, 0.86 mL / g, 1.2 mL / g, 1.33 mL / g, 1.39 mL / g, 1.42 mL / g, 1.5 mL / g, 1.6 mL / g, 1.7 mL / g, or 1.78 mL / g; In the present invention, the density of the epoxy ligand in the chromatography material is preferably 35 to 60 μmol / mL, for example, 39 μmol / mL, 42 μmol / mL, 43 μmol / mL, 48 μmol / mL, or 51 μmol / mL.

[0058] The present invention further provides a method for preparing a chromatography material, the steps of which include pre-polymerizing the aforementioned chromatography material raw material composition and then molding it to complete the process, The temperature of the prepolymerization reaction is 0°C to 10°C, The molding temperature is 25°C to 60°C.

[0059] In the present invention, the temperature of the prepolymerization reaction is preferably 2°C to 8°C.

[0060] In the present invention, the time for the prepolymerization reaction may be 15 to 60 minutes, preferably 20 to 40 minutes, for example, 30 minutes.

[0061] In the present invention, the molding temperature is preferably 30°C to 40°C.

[0062] In the present invention, the molding time may be 2 hours to 12 hours, and is preferably 4 hours to 8 hours.

[0063] In the present invention, the molding process is maintained in a stationary state and is not affected by external forces.

[0064] In the present invention, the mold may be a mold that is common in the art, and the material of the mold may be a material that is common in the art, such as PE plastic, and the shape of the mold may be a shape that is common in the art, such as a cylindrical shape.

[0065] In the present invention, after the shaping is completed, the method preferably further comprises post-treatment of the chromatographic material.

[0066] Here, the post-treatment method may be a method commonly used in the art, and for example, the post-treatment may involve washing the chromatography material with an alcohol organic reagent.

[0067] Here, the alcohol organic reagent may be any conventional one in the art, such as one or more of anhydrous methanol, ethanol and isopropanol.

[0068] Here, the cleaning method may be a conventional method in the field, such as vacuum cleaning or suction filtration and leaching.

[0069] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; In the chromatography material raw material composition, the raw material A is a polyepoxy glycidyl ether polymer and a monoepoxy glycidyl ether monomer, and the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide (DMF), dimethyl sulfoxide (DMSO), and methyl t-butyl ether.

[0070] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; In the chromatography material raw material composition, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide (DMF), dimethyl sulfoxide (DMSO), and methyl t-butyl ether, and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether.

[0071] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; In the chromatography material raw material composition, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, the porogen B is one or more of toluene, dioxane, and methyl t-butyl glyceryl ether, and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether.

[0072] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; The chromatography material raw material composition contains 50 to 70% raw material A, where the total weight of raw material A and porogen B is 100%, and the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether monomer, and the monoepoxy glycidyl ether monomer is phenyl glycidyl ether and / or butyl glycidyl ether, and 30 to 50% porogen B, where the porogen B forms "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane".

[0073] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; The chromatography material raw material composition contains 50 to 70% raw material A, where the total weight of raw material A and porogen B is 100%, and the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether monomer, and the monoepoxy glycidyl ether monomer is phenyl glycidyl ether and / or butyl glycidyl ether, and 30 to 50% porogen B, where the porogen B is "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane".

[0074] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; The chromatography material raw material composition contains the following components: 50 to 70% raw material A, where the total weight of raw material A and porogen B is 100%, the raw material A being polyglycerol triglycidyl ether and a monoepoxy glycidyl ether monomer, the monoepoxy glycidyl ether monomer being phenyl glycidyl ether and / or butyl glycidyl ether; 30 to 50% porogen B, where the porogen B is "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane"; and 1‰ of catalyst C.

[0075] In some preferred embodiments of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C, and then completing the process; The chromatography material raw material composition contains the following components, where the total weight of raw material A and porogen B is 100%: 60% raw material A, which is polyglycerol triglycidyl ether having a molecular weight of 550 and butyl glycidyl ether, wherein the volume ratio of the polyglycerol triglycidyl ether having a molecular weight of 550 to the butyl glycidyl ether is 2:1; 40% porogen B, which is dioxane and / or methyl t-butyl glyceryl ether, wherein the volume ratio of the dioxane to the methyl t-butyl glyceryl ether is 1:1; and 1‰ of catalyst C.

[0076] In one preferred embodiment of the present invention, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 4°C, followed by molding at 35°C, to complete the process; The chromatography material raw material composition contains the following components, where the total weight of raw material A and porogen B is 100%: 60% raw material A, which is polyglycerol triglycidyl ether having a molecular weight of 550 and butyl glycidyl ether, where the volume ratio of the polyglycerol triglycidyl ether having a molecular weight of 550 to the butyl glycidyl ether is 2:1; 40% porogen B, which is dioxane and methyl t-butyl glyceryl ether, where the volume ratio of the dioxane to the methyl t-butyl glyceryl ether is 1:1; and 1‰ of catalyst C, which is boron trifluoride diethyl ether.

[0077] The present invention further provides a chromatographic material prepared by the above-described preparation method.

[0078] The present invention further provides a monolithic column comprising the aforementioned chromatographic material.

[0079] The present invention further provides a monolithic column having through-holes with an average pore size of 0.2 to 10 μm distributed inside.

[0080] The porosity of the through holes is 30 to 80%, the pore volume of the through holes is 0.5 to 2.5 mL / g; The density of the epoxy ligand in the monolith column is 30 to 80 μmol / mL.

[0081] In the monolith column of the present invention, the average pore size of the through holes is preferably 0.4 to 8 μm, for example, 0.44 μm, 1.4 μm, 0.74 μm, 0.8 μm, 2 μm, 2.7 μm, 3.4 μm, 3.7 μm, 3.8 μm, 4.2 μm, 4.3 μm, 4.5 μm, 5.2 μm, 6 μm, 7 μm, or 7.5 μm.

[0082] In the monolith column of the present invention, the porosity of the through holes is preferably 40 to 68%, for example, 45.9%, 46.5%, 46.8%, 48.8%, 49.7%, 59.7%, 58.6%, 60.1%, 60.4%, 65.1%, 65.9%, or 66.3%.

[0083] In the monolith column of the present invention, the pore volume of the through holes is preferably 0.5 to 1.8 mL / g, for example, 0.53 mL / g, 0.58 mL / g, 0.59 mL / g, 0.62 mL / g, 0.86 mL / g, 1.2 mL / g, 1.33 mL / g, 1.39 mL / g, 1.42 mL / g, 1.5 mL / g, 1.6 mL / g, 1.7 mL / g, or 1.78 mL / g; In the present invention, the density of the epoxy ligand in the monolith column is preferably 35 to 60 μmol / mL, for example, 39 μmol / mL, 42 μmol / mL, 43 μmol / mL, 48 μmol / mL, or 51 μmol / mL.

[0084] The present invention further provides a chromatography stack comprising the aforementioned raw material composition, the aforementioned chromatography material, or the aforementioned monolith column.

[0085] The chromatographic stack in the present invention refers to a chromatographic separation material, such as a polyepoxy group or polyhydroxy group-containing chromatographic material.

[0086] The present invention further provides an application of the aforementioned chromatography material raw composition, or the aforementioned chromatography material, or the aforementioned monolith column, or the aforementioned chromatography stack in the field of biopolymer purification.

[0087] In the present invention, the biopolymer is generally a protein, a plasmid DNA, an mRNA, a viral vector, an exosome, an ADC, or a complex macromolecular protein.

[0088] The above preferred conditions can be arbitrarily combined in accordance with common knowledge in the art to obtain preferred embodiments of the present invention.

[0089] All of the reagents and raw materials used in the present invention are commercially available. [Effects of the Invention]

[0090] The positive effects of the present invention are as follows:

[0091] 1. The raw materials contained in the chromatography material composition of the present invention can be used in combination with porogens and catalysts to prepare chromatography materials with controllable through-pore (submicron to micron) structures, and can be grafted with different ligand groups through certain chemical coupling modifications to achieve chromatography functions. The advantages of this method are that it can solve the current problems associated with the application of chromatography microsphere fillers in biopolymers, specifically:

[0092] (1) The pores of the chromatographic materials prepared by the present invention are tunable, submicron-micron order, relatively uniform in distribution, exhibiting a unimodal normal distribution, and have high ligand utilization.

[0093] (2) The pores of the chromatography material prepared by the present invention are through-holes, which have no diffusion effect, do not require an increase in retention time, shorten the process cycle, have no significant eddy current phenomenon, have low shear force, and have a high effective activity yield of biopolymers.

[0094] (3) The chromatographic materials prepared in this invention do not affect sample loading at high flow rates.

[0095] (4) The preparation process of the chromatography material of the present invention is relatively simple, batch-to-batch stability is easily achieved, and column packing verification work is not required, and it can be used directly (plug-and-play type).

[0096] (5) The chromatographic materials prepared by the present invention can be reused through batch-to-batch verification and can also be used as disposable technical consumables, with high economic selectivity and low production costs.

[0097] 2. The chromatography material of the present invention has the characteristics of a uniform internal structure, high preparation reproducibility, high toughness, the ability to be modified with functional ligands, and relatively high hydrophilicity, making it more suitable for the separation and purification of biomolecules. [Brief explanation of the drawings]

[0098] [Figure 1] 1A is a schematic cross-sectional view of a molding jig for a chromatography material, and FIG. 1B is a schematic longitudinal-sectional view of a molding jig for a chromatography material. [Figure 2] FIG. 1 is a scanning electron microscope image of the chromatographic material prepared in Example 1. [Figure 3] FIG. 1 is a scanning electron micrograph of the chromatographic material prepared in Example 3. [Figure 4] FIG. 1 is a scanning electron micrograph of the chromatographic material prepared in Example 6. [Figure 5]FIG. 1 is a schematic diagram of the pore size distribution of the chromatographic material prepared in Example 2. [Figure 6] FIG. 1 is a schematic diagram of the pore size distribution of the chromatographic material prepared in Example 7. [Figure 7] FIG. 1 is a schematic diagram of the bovine serum albumin loading under different flow rates of the chromatographic material prepared in Example 6. DETAILED DESCRIPTION OF THE INVENTION

[0099] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the examples. In the following examples, experimental methods for which specific conditions are not specified are selected according to conventional methods and conditions or product instructions.

[0100] Unless otherwise specified, the reagents used in the following examples and comparative examples may be any reagents that are commonly available commercially in the art.

[0101] Example 1 1. 30% by weight of raw material A (polyglycerol glycidyl ether, Mr=550), 70% by weight of porogen B (toluene and dioxane in a 1:1 volume ratio), and 1‰ of catalyst C (boron trifluoride) were accurately weighed and mixed thoroughly. The mixture was stirred at 4°C for 30 minutes to obtain a prepolymer. 2. The uniformly stirred prepolymer is poured into a PE plastic material mold (a self-made mold, the shape of which is a concentric circular cylinder, the structure diagram of which is shown in Figure 1), and then left to stand for 4 hours, the temperature of which is controlled at 35°C, and the molding process is kept statically stable. 3. After the reaction is completed, add anhydrous methanol or alcoholic organic reagent and then vacuum wash or suction filter to remove the remaining liquid in the pores. 4. After cleaning was completed, the specimen was immersed in a 20% ethanol solution and kept at a low temperature (4°C) for use.

[0102] In Examples 2 to 7, the conditions were all the same as in Example 1, except for the conditions shown in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0103] Note: A%, B%, and C% in Table 1 represent the weight percentages of raw material A, porogen B, and catalyst C, respectively, in the total amount of "A+B."

[0104] Example of effect

[0105] (1) Field emission scanning electron microscope test Test subjects: chromatographic materials prepared in Examples 1, 3, and 6. Testing methods: The testing methods were conventional methods in the field. Test equipment: Germany-Zeiss Sigma300, Oxford Spectrum. Test results: As shown in Figure 2, Figures 2 to 4 correspond to scanning electron microscope images measured by the chromatography materials prepared in Examples 1, 3, and 6, respectively. Figures 2 to 4 show that the chromatography materials prepared in Examples 1, 3, and 6 all have pore structures, where Figure 2 shows that the pores of the chromatography material prepared in Example 1 are the largest, and Figure 4 shows that the pores of the chromatography material prepared in Example 6 are the smallest. Thus, the proportion of raw material A affects the pore size, and the pore size can be adjusted by controlling the proportion of raw material A.

[0106] (2) Pore size distribution, pore volume, and porosity tests Test subjects: chromatographic materials prepared in Examples 1 to 13 and Comparative Examples 1 to 10. Test method: Mercury porosimetry. Test equipment: American McMurrittic autoporeV 9620.

[0107] Specific test operations: A certain amount of chromatographic material was placed in absolute ethanol and subjected to a vacuum for 30 minutes to displace the solvent in the porous material. The absolute ethanol on the surface was then dried, and the sample was then placed in an oven at 50-60°C for at least 2 hours. After drying, the sample was placed on the sample stage of the mercury intrusion apparatus and mercury was injected into the apparatus. The test parameters were as follows: Pressure range: 0.10 to 61,000.00 psia Sample mass: 0.2104g Capillary utilization rate: 28% Mercury temperature: 15.75℃ Total mass: 111.1832g Penetrometer volume: 4.2065 mL Mass of the penetrometer: 61.1553g Contact angle: 130.000° Penetrometer ID: 15-0295-(15) 3 Bulb, 1.190 Stem, Solid

[0108] Test Results: The pore uniformity throughout the chromatography materials of Examples 2 and 7 was intuitively observed to be a single-peak normal pore distribution as shown in Figures 5-6 (Figures 5-6 are the original instrumental test diagrams, and the data in the figures are the original test data; the ordinate represents the logarithm of the differential mercury intrusion volume, and the abscissa represents the pore size converted through instrumental analysis). Figures 5-6 show that the pore size distributions of the chromatography materials prepared in Examples 2 and 7 exhibit a good normal distribution without obvious bimodalities (i.e., no double pore size distribution phenomenon). The average pore size results were consistent with the results obtained by electron microscopy. With increasing addition of raw material A, the pore size of the prepared chromatography materials decreased. The average pore size of the chromatography material prepared in Example 2 was 4.2 μm, which was a large pore on the micron order, while the average pore size of the chromatography material prepared in Example 7 was 0.74 μm, which was a large pore on the submicron order.

[0109] Specific data measured in Examples 1 to 13 and Comparative Examples 1 to 6 are shown in Table 2 below. [Table 2]

[0110] Note: The average pore diameter was positively correlated with the porosity and pore volume. The optimum range for pore volume was 0.5-1.8 mL / g, the optimum range for average pore diameter was 0.4-6 μm, and the optimum range for porosity was 40-68%.

[0111] As can be seen from Table 2, the pore volumes of the chromatography materials prepared in Examples 1 to 13 were generally 0.53 to 1.7 mL / g, the average pore diameters were 0.51 to 5.2 μm, and the porosities were 45.9 to 67.1%. The pore volumes, average pore diameters, and porosities of the chromatography materials prepared in Examples 1 to 13 were varied by adjusting the type of raw material A, the type of porogen B, and the relative ratios of raw material A and porogen B. The pore volumes, average pore diameters, and porosities of Examples 6 and 7 were almost the same, with little difference.

[0112] Compared with Example 1, the content of raw material A in Comparative Example 1 was relatively high, and the prepared chromatographic material was without pores and could not meet the application needs.

[0113] Compared with Example 2, the content of catalyst C in Comparative Example 2 was relatively high, and the prepared chromatographic material was without pores and could not meet the application needs.

[0114] In Comparative Examples 3 and 4, raw material A consisted solely of monoepoxy glycidyl ether monomers (butyl glycidyl ether in Comparative Example 3, and phenyl glycidyl ether in Comparative Example 4), and the prepared chromatography materials were viscous and could not meet the application needs.

[0115] Compared with Example 6, in Comparative Example 5, the temperature of the prepolymerization reaction was relatively low, and the prepared chromatographic material cracked, which could not meet the application needs.

[0116] Compared with Example 6, the temperature of the prepolymerization reaction in Comparative Example 6 was relatively high, and the prepared chromatographic material fell off, which could not meet the application needs.

[0117] Compared with Example 6, in Comparative Example 7, the prepolymerization reaction and molding temperatures were the same, both 25°C, and the prepared chromatographic material fell off, which could not meet the application needs.

[0118] Compared with Example 6, in Comparative Example 8, the prepolymerization reaction and molding temperatures were the same, both 5°C, and the prepared chromatographic material was in a viscous state, which could not meet the application needs.

[0119] Compared with Example 6, in Comparative Example 9, the molding temperature was relatively low, and the prepared chromatographic material fell off, which could not meet the application needs.

[0120] Compared with Example 6, in Comparative Example 10, the molding temperature was relatively high, and the prepared chromatographic material was without pores and could not meet the application needs.

[0121] (3) Epoxy group ligand density test Test subjects: chromatographic materials prepared in Examples 1 to 13 and Comparative Examples 1 to 10 modified with epichlorohydrin.

[0122] Because the amount of epoxy ligand contained in the chromatographic material itself is relatively small, direct measurement cannot accurately quantify it, so the chromatographic material had to be first "epichlorohydrin modified."

[0123] "Epichlorohydrin modification" involved pretreating the chromatography materials of Examples 1 to 13 and Comparative Examples 1 to 10 with epichlorohydrin before conducting the epoxy group ligand density test. The reagents used in this pretreatment and their amounts were 10 wt% epichlorohydrin, 44 wt% deionized water, 44 wt% dimethyl sulfoxide (DMSO), and 2 wt% sodium hydroxide, respectively. The percentages refer to the percentages of each reagent based on the total weight of the reagents. The mass ratio of the chromatography material to epichlorohydrin was 1:1. The pretreatment reaction temperature was 45°C, and the reaction time was 10 hours. Apart from the conditions specifically described above, other conditions may be those commonly used in the art for this pretreatment.

[0124] Test method: Sodium thiosulfate-hydrochloric acid titration method.

[0125] The epichlorohydrin-modified chromatography materials of Examples 1-13 and Comparative Examples 1-10 were each washed with deionized water, placed in a sand-core funnel, and vacuum-dried for 10 minutes. One piece (approximately 0.5 g) was then weighed and placed in a polished-nosed Erlenmeyer flask. 3 mL of 1.3 mol / L sodium thiosulfate and 1-2 drops of phenolphthalein indicator were added, the Erlenmeyer flask was sealed, and the mixture was allowed to react at room temperature for 1 hour. The supernatant was then titrated with 0.1 mol / L hydrochloric acid standard solution until the red solution became colorless. Based on the volume of hydrochloric acid standard solution consumed, the density of the epoxy group ligand was calculated using the following equation: S=M HCl (V0-V1)*ρ / W S: density of epoxy group ligand, mol / L; M HCl : Hydrochloric acid concentration, mol / L, V0, V1: Volume of HCl before and after titration, mL; ρ: medium density (1.2g / mL), W: The measured mass of the entire block. Test equipment: Acid-base titrator. Test results: shown in Table 3 below. [Table 3]

[0126] Note: The " / " in Table 3 indicates that the density data of the epoxy ligand was not measured because the structure of the chromatographic material did not meet the requirements (e.g., the chromatographic material had no through-holes; the chromatographic material was in a viscous state; the chromatographic material had fallen off and slugs had fallen; the chromatographic material had no holes, etc.).

[0127] As shown in the data in Table 3 above, after modification using the conventional epoxidation modification method of epichlorohydrin, the epoxy group density on the prepared chromatography material was a total of 35-60 μmol / mL, which corresponds to the epoxy group density of the chromatography filler microspheres and can meet the requirements for chromatographic separation and analysis. Furthermore, the magnitude of the epoxy group density was positively correlated with the amount of raw material A used in the preparation process of the chromatography material and the epoxy value of the polymer or monomer in the raw material A used. The epoxy group ligand density of the chromatography materials obtained in Examples 1-13 could be changed by adjusting conditions such as the type of raw material A, the type of porogen B, the relative proportions of raw material A and porogen B used, and the amount of catalyst used. Here, the epoxy group ligand densities of Examples 6-7 were almost the same with little difference. Specifically, Compared with Example 1, the content of raw material A in Comparative Example 1 was relatively high, and the density of epoxy group ligands in the prepared chromatography material was only 6 μmol / mL, which could not meet the application needs.

[0128] Compared with Example 2, the content of catalyst C in Comparative Example 2 was relatively high, and the density of epoxy group ligands in the prepared chromatography material was only 8 μmol / mL, which could not meet the application needs.

[0129] In Comparative Examples 3 and 4, raw material A consisted solely of monoepoxy glycidyl ether monomers (butyl glycidyl ether in Comparative Example 3, and phenyl glycidyl ether in Comparative Example 4), and the prepared chromatography materials were in a viscous state, making it impossible to measure the density data of the epoxy group ligands.

[0130] Compared with Example 6, in Comparative Example 5, the temperature of the prepolymerization reaction was relatively low, and the prepared chromatographic material was cracked, so the density data of the epoxy group ligand could not be measured.

[0131] Compared with Example 6, in Comparative Example 6, the temperature of the prepolymerization reaction was relatively high, and the prepared chromatographic material fell off, so that the density data of the epoxy group ligand could not be measured.

[0132] Compared with Example 6, in Comparative Example 7, the prepolymerization reaction and molding temperatures were the same, both at 25°C, and the prepared chromatography material fell off, making it impossible to measure the density data of the epoxy group ligand.

[0133] Compared with Example 6, in Comparative Example 8, the prepolymerization reaction and molding temperatures were the same, both at 5°C. The prepared chromatography material was in a viscous state, and the density data of the epoxy group ligand could not be measured.

[0134] Compared with Example 6, in Comparative Example 9, the molding temperature was relatively low, and the prepared chromatography material fell off, so the density data of the epoxy group ligand could not be measured.

[0135] Compared with Example 6, in Comparative Example 10, the molding temperature was relatively high, and the prepared chromatography material was non-porous, and the density data of its epoxy group ligand could not be measured.

[0136] (4) Biopolymer yield test Test subjects: chromatographic materials prepared in Examples 6 and 7.

[0137] Test method: Chromatography equipment and chromatography column (BIA 1 mL monolith column fixture was used to incorporate the chromatography materials from Examples 6 and 7) were used to measure the protein content by the Lorry method (experimental method for protein content measurement using the Filin-Phenol reagent method). The hydrophobicity of the chromatography materials from Examples 6 and 7 and the yield of bovine serum albumin after chromatographic binding and elution were measured. The chromatographic mobile phases were equilibration solution A: phosphate buffer PB (pH = 6.5) + 0.8 M ammonium sulfate, and elution solution B: phosphate buffer PB (pH = 6.5). The chromatography flow rate was 1 mL / min. After equilibration with 10 mL of equilibration solution A, 30 mL of sample (bovine serum albumin dissolved in equilibration solution, concentration: 2 mg / mL) was injected. The chromatography column was washed with 5-10 mL of equilibration solution, and then eluted with elution solution B. The volume was 3-4 mL. The eluate was collected and the protein concentration was measured.

[0138] Test equipment: Akata explore 100 chromatography equipment.

[0139] Test results are shown in Table 4 below. [Table 4]

[0140] As can be seen from the data in Table 4, the bovine serum albumin chromatographic yields of the chromatographic materials prepared in Examples 6 and 7 were 90% or higher (generally, a yield of 90% or higher is desirable for single-step chromatography). The bovine serum albumin yield of the chromatographic material prepared in Example 6 was 95%, and the bovine serum albumin yield of the chromatographic material prepared in Example 7 was 98%. This may be due to the stronger hydrophobic effect of phenyl than that of butyl, i.e., the stronger binding effect of phenyl in the same mobile phase, the larger elution volume, and the stronger elution strength, resulting in a higher yield of butyl under the same eluent conditions. The nonspecific irreversible adsorption of the hydrophobic chromatographic material prepared in Example 7 was shown to be lower than that of the chromatographic material prepared in Example 6.

[0141] (5) Sample loading test under different flow rates Test subject: chromatographic material prepared in Example 6.

[0142] Test method: The protein content was measured using a chromatography device and a chromatography column (a 1 mL monolith column fixture manufactured by BIA was used and the chromatography material prepared in Example 6 was incorporated and tested) by the Lorry method. The amount of bovine serum albumin adsorbed (i.e., the loading amount) by the chromatography material prepared in Example 6 at different flow rates was measured. The mobile phase and chromatography method were the same as those used in (4) Biopolymer Yield Test.

[0143] Test equipment: Akata explore 100 chromatography equipment.

[0144] The test results are shown in Table 5 and Figure 7. [Table 5]

[0145] Note: DBC was the dynamic binding load of bovine serum albumin on the chromatographic material.

[0146] As can be seen from Table 5 and Figure 7, increasing the flow rate had little effect on protein adsorption. Conventional chromatography microsphere media generally require a high flow rate within a retention time of 2 min (120 s) and generally require retention for 3 to 4 min or more. However, the retention time of the chromatography material prepared in Example 6 of the present invention was shorter, and as the flow rate increased, the loading capacity of the conventional chromatography microsphere filler decreased by twofold, while the change in loading capacity of the chromatography material prepared in Example 6 of the present invention was smaller.

[0147] (6) Lot-to-lot stability test Test subject: chromatographic material prepared in Example 7.

[0148] Test method: Chromatography equipment and chromatography column (BIA 1 mL monolith column fixture, inner device incorporating the chromatographic material prepared in Example 7) were used to test the batch-to-batch stability of the chromatography process. The test sample was a mixture of two proteins: bovine serum albumin (Yancheng Saibao Biotechnology Co., Ltd.) and human immunoglobulin IgG (Guangdong Shuanglin Biopharmaceutical Co., Ltd.) (prepared with equilibrium solution A above, with the final concentration of both proteins at 2 mg / mL). The mobile phase and chromatography method were the same as those used in (4) Biopolymer Yield Test.

[0149] Test equipment: Akata explore 100 chromatography equipment.

[0150] The test results are shown in Table 6. [Table 6]

[0151] The data in Table 6 provide an intuitive understanding of the batch-to-batch stability of the chromatography process. The normal distribution curves were essentially consistent, with relatively high reproducibility. The peak appearing within the retention time range of 0 to 8 min was characteristic of bovine serum albumin, while the peak appearing within the retention time range of 8 to 13.5 min was characteristic of human immunoglobulin IgG. Human immunoglobulin IgG is more hydrophobic than bovine serum albumin, resulting in delayed peak appearance. The overlap in peak appearance time and size between batches was relatively good (see Table 6 for details). The slight deviations in the data in Table 6 were due to manual sample injection during the production process; the use of automated sample injection and chromatography during production eliminated this error.

Claims

1. A chromatography material raw material composition, the total weight of raw material A and porogen B being 100%, 20% to 70% of raw material A, 30% to 80% of Porogen B; 0.1‰ to 1‰ of catalyst C; The chromatography material raw material composition, wherein the raw material A contains a polyepoxy group-containing substance.

2. One or more of the following conditions a to k are satisfied: a. The polyepoxy group-containing substance is a polyepoxy glycidyl ether-based substance or a polyepoxy glycidyl ester-based substance; The glycidyl ether-based material is preferably one or more of the monomers satisfying the following structural formula I: 【Chemistry 1】 In structural formula I, R １ represents a hydrogen atom, a substituted or unsubstituted C １ ~C １０ or an epoxy group, n is a positive integer from 0 to 10, L １ is selected from an oxygen atom or a nitrogen atom, L ２ is selected from an oxygen atom or a nitrogen atom, X is a substituted or unsubstituted C １ ~C １０ an alkylene group represented by the formula (I), a substituted or unsubstituted C ３ ~C １０ a cycloalkylene group of the formula: 【Chemistry 2】 or a benzene ring; The glycidyl ether-based substance is preferably one or more of the following monomers (a) to (m): 【Transformation 3】 The glycidyl ether-based material is preferably one or more of the polymers satisfying the following structural formula II: 【Chemistry 4】 In structural formula II, R ２ represents a hydrogen atom or a substituted or unsubstituted C １ ~C １０ or an alkyl group of 【Transformation 5】 is selected from m is a positive integer from 2 to 40; The glycidyl ether-based substance is preferably one or more of the following polymers (1) to (4): 【Transformation 6】 The glycidyl ester-based material is preferably one or more of the monomers satisfying structural formula III: 【Transformation 7】 In Formula III, Y is a substituted or unsubstituted C １ ~C １０ an alkylene group represented by the formula (I), a substituted or unsubstituted C ３ ~C １０ or a benzene ring, The glycidyl ester-based substance is preferably the following monomer (A) and / or (B): 【Transformation 8】 b. The polyepoxy group-containing substance is one or more of the following monomers (I) to (IV): 【Chemistry 9】 c. The polyepoxy group-containing substance is a polyepoxy glycidyl ether-based monomer and / or a polyepoxy glycidyl ether-based polymer, d. The molecular weight of the polyepoxy group-containing substance is 300 to 2000, for example, 550 or 700; e. the percentage by weight of the ingredient A relative to the total weight of the ingredient A and the porogen B is 22% to 70%, for example, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 58%, 65%, or 68%; f. The raw material A may further contain a monoepoxy group-containing substance, The monoepoxy group-containing substance is preferably a monoepoxy glycidyl ether-based substance or a monoepoxy glycidyl ester-based substance, g. the porogen B is one or more chemical solvents compatible with the feedstock A and the catalyst C, preferably one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide, dimethyl sulfoxide, and methyl t-butyl ether, more preferably one or more of toluene, dioxane, and methyl t-butyl ether, such as toluene and dioxane, toluene and methyl t-butyl ether, or dioxane and methyl t-butyl ether; h. When the porogen B is a mixture of two different substances, the volume ratio of the two different substances is preferably (0.1-9):1, for example, 0.5:1, 1:1, 2:1, 3:1, 5:1, or 7:1; i. the percentage by weight of the porogen B relative to the total weight of the raw material A and the porogen B is preferably 35% to 80%, for example, 40%, 45%, 50%, 60%, 65%, 70%, 75%, or 78%; j. The catalyst C is a Lewis acid and / or a Lewis acid complex; k. The chromatography material raw material composition of claim 1, wherein the weight percentage of the catalyst C relative to the total weight of the raw material A and the porogen B is preferably 0.3‰ to 1‰, for example, 0.4‰, 0.5‰, 0.6‰, 0.7‰, 0.8‰, or 0.9‰.

3. One or more of the following conditions a to g are satisfied: a. The polyepoxy glycidyl ether monomer is glycerol triglycidyl ether and / or pentaerythritol tetraglycidyl ether, b. The polyepoxy glycidyl ether-based polymer is polyglycerol glycidyl ether and / or polypentaerythritol tetraglycidyl ether; c. When the polyepoxy group-containing substance is a mixture of the polyepoxy glycidyl ether monomer and the polyepoxy glycidyl ether polymer, the mixing volume ratio of the polyepoxy glycidyl ether monomer to the polyepoxy glycidyl ether polymer is (0.1 to 9):1, for example, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, or 8:1; d. The monoepoxy glycidyl ether-based substance is phenyl glycidyl ether and / or butyl glycidyl ether; e. the monoepoxy glycidyl ester-based material is one or more of phenyl glycidyl ether, butyl glycidyl ether, pentyl glycidyl ether, octyl glycidyl ether, octadecyl glycidyl ether, and naphthyl glycidyl ether, preferably phenyl glycidyl ether and / or butyl glycidyl ether; f. the Lewis acid is one or more of aluminum trichloride, boron trifluoride, iron bromide, iron chloride, zinc chloride, niobium trichloride or niobium pentachloride, and sulfur trioxide, for example, boron trifluoride; g. the Lewis acid complex is one or more of boron trifluoride diethyl etherate, boron trifluoride acetonitrile, boron trifluoride dimethylcarbonate, and boron trifluoride ethylamine; Preferably, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide, dimethyl sulfoxide, and methyl t-butyl ether, and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether; Preferably, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, the porogen B is one or more of toluene, dioxane, and methyl t-butyl glyceryl ether, and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether; Preferably, the chromatography material raw material composition comprises 50 to 70% raw material A, where the total weight of raw material A and porogen B is 100%, the raw material A being polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, and the monoepoxy glycidyl ether-based monomer being phenyl glycidyl ether and / or butyl glycidyl ether, and 30 to 50% porogen B, where the porogen B is "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane", 3. The chromatography material raw material composition according to claim 1, wherein the total weight of raw material A and porogen B is taken as 100%, and the chromatography material raw material composition preferably comprises: 60% raw material A, which is polyglycerol triglycidyl ether having a molecular weight of 550 and butyl glycidyl ether, wherein the volume ratio of the polyglycerol triglycidyl ether having a molecular weight of 550 to the butyl glycidyl ether is 2:1; 40% porogen B, which is dioxane and methyl t-butyl glyceryl ether, wherein the volume ratio of the dioxane to the methyl t-butyl glyceryl ether is 1:1; and 1% of catalyst C, which is boron trifluoride diethyl ether.

4. A chromatographic material comprising the starting composition according to at least one of claims 1 to 3, the chromatography material has through-holes distributed therein with an average pore size of 0.2 to 10 μm; the porosity of the through holes is 30 to 80%; the through holes have a pore volume of 0.5 to 2.5 mL / g; The chromatography material has a density of the epoxy ligand of 30 to 80 μmol / mL.

5. One or more of the following conditions a to d are satisfied: a. The average pore diameter of the through holes is 0.4 to 8 μm, for example, 0.44 μm, 1.4 μm, 0.74 μm, 0.8 μm, 2 μm, 2.7 μm, 3.4 μm, 3.7 μm, 3.8 μm, 4.2 μm, 4.3 μm, 4.5 μm, 5.2 μm, 6 μm, 7 μm, or 7.5 μm; b. The porosity of the through holes is 40 to 68%, for example, 45.9%, 46.5%, 46.8%, 48.8%, 49.7%, 59.7%, 58.6%, 60.1%, 60.4%, 65.1%, 65.9%, or 66.3%; c) the pore volume of the through-holes is 0.5 to 1.8 mL / g, for example, 0.53 mL / g, 0.58 mL / g, 0.59 mL / g, 0.62 mL / g, 0.86 mL / g, 1.2 mL / g, 1.33 mL / g, 1.39 mL / g, 1.42 mL / g, 1.5 mL / g, 1.6 mL / g, 1.7 mL / g, or 1.78 mL / g; d. the density of the epoxy ligand in the chromatographic material is 35-60 μmol / mL, for example, 39 μmol / mL, 42 μmol / mL, 43 μmol / mL, 48 μmol / mL, or 51 μmol / mL; The chromatography material according to claim 4, wherein the chromatography material preferably satisfies conditions a to d simultaneously.

6. A method for preparing a chromatography material, the steps of which include pre-polymerizing the chromatography material raw material composition according to at least one of claims 1 to 3 and then shaping the composition, and completing the process; The temperature of the prepolymerization reaction is 0°C to 10°C, The molding temperature is 25°C to 60°C, The temperature of the prepolymerization reaction is preferably 2°C to 8°C, The time for the prepolymerization reaction is preferably 15 to 60 minutes, more preferably 20 to 40 minutes, for example, 30 minutes. The molding temperature is preferably 30°C to 40°C, The molding time is preferably 2 hours to 12 hours, more preferably 4 hours to 8 hours, Preferably, the molding process is performed while the molded product is kept stationary and is not affected by external forces. Preferably, in the method for preparing a chromatography material, the steps include pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C to complete the process; in the chromatography material raw material composition, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer; the porogen B is one or more of toluene, xylene, dichloromethane, dichloroethane, dioxane, formamide, dimethyl sulfoxide, and methyl t-butyl ether; and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether; Preferably, in the method for preparing a chromatography material, the steps include pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, and then molding the composition at 30°C to 40°C to complete the process; in the chromatography material raw material composition, the raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer; the porogen B is one or more of toluene, dioxane, and methyl t-butyl glyceryl ether; and the monoepoxy glycidyl ether-based monomer is preferably phenyl glycidyl ether and / or butyl glycidyl ether; Preferably, the steps of the method for preparing a chromatography material include pre-polymerizing a chromatography material raw material composition at 2°C to 8°C, followed by molding at 30°C to 40°C to complete the process, and the chromatography material raw material composition comprises 50 to 70% of raw material A, where raw material A is polyglycerol triglycidyl ether and a monoepoxy glycidyl ether-based monomer, and the monoepoxy glycidyl ether-based monomer is phenyl glycidyl ether and / or butyl glycidyl ether, and 30 to 50% of porogen B, where porogen B is "dioxane and methyl t-butyl glyceryl ether" or "toluene and dioxane," where the total weight of raw material A and porogen B is 100%. Preferably, the method for preparing a chromatography material includes the steps of pre-polymerizing a chromatography material raw material composition at 2°C to 8°C and then molding the composition at 30°C to 40°C to complete the process, and the chromatography material raw material composition comprises the following components, where the total weight of raw material A and porogen B is 100%, the chromatography material raw material composition includes: 60% raw material A, which is polyglycerol triglycidyl ether having a molecular weight of 550 and butyl glycidyl ether, and the volume ratio of the polyglycerol triglycidyl ether having a molecular weight of 550 to the butyl glycidyl ether is 2:1; 40% porogen B, which is dioxane and methyl t-butyl glyceryl ether, and the volume ratio of the dioxane to the methyl t-butyl glyceryl ether is 1:1; and 1% of catalyst C, which is boron trifluoride diethyl ether.

7. A chromatographic material prepared by the method of claim 6.

8. A monolithic column, characterized in that it comprises a raw material composition according to at least one of claims 1 to 3, or a chromatography material according to at least one of claims 4 to 5 and 7.

9. A monolithic column, wherein through-holes having an average pore diameter of 0.2 to 10 μm are distributed inside the monolithic column; the porosity of the through holes is 30 to 80%; the through holes have a pore volume of 0.5 to 2.5 mL / g; The density of the epoxy ligand in the monolith column is 30 to 80 μmol / mL.

10. One or more of the following conditions a to d are satisfied: a. The average pore diameter of the through holes is 0.4 to 8 μm, for example, 0.44 μm, 1.4 μm, 0.74 μm, 0.8 μm, 2 μm, 2.7 μm, 3.4 μm, 3.7 μm, 3.8 μm, 4.2 μm, 4.3 μm, 4.5 μm, 5.2 μm, 6 μm, 7 μm, or 7.5 μm; b. The porosity of the through holes is 40 to 68%, for example, 45.9%, 46.5%, 46.8%, 48.8%, 49.7%, 59.7%, 58.6%, 60.1%, 60.4%, 65.1%, 65.9%, or 66.3%; c) the pore volume of the through-holes is 0.5 to 1.8 mL / g, for example, 0.53 mL / g, 0.58 mL / g, 0.59 mL / g, 0.62 mL / g, 0.86 mL / g, 1.2 mL / g, 1.33 mL / g, 1.39 mL / g, 1.42 mL / g, 1.5 mL / g, 1.6 mL / g, 1.7 mL / g, or 1.78 mL / g; d. The density of the epoxy ligand in the monolith column is 35 to 60 μmol / mL, for example, 39 μmol / mL, 42 μmol / mL, 43 μmol / mL, 48 μmol / mL, or 51 μmol / mL; The monolithic column according to claim 9, wherein the monolithic column preferably satisfies conditions a to d simultaneously.

11. A chromatography stack comprising a raw material composition according to at least one of claims 1 to 3, a chromatography material according to at least one of claims 4 to 5 and 7, or a monolithic column according to at least one of claims 8 to 10.

12. Application of the raw material composition according to at least one of claims 1 to 3, or the chromatography material according to at least one of claims 4 to 5 and 7, or the monolith column according to at least one of claims 8 to 10 in the field of biopolymer purification.

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