CN114805861A - Preparation method of polystyrene microspheres for hydrophilic biological separation - Google Patents
Preparation method of polystyrene microspheres for hydrophilic biological separation Download PDFInfo
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- 239000004005 microsphere Substances 0.000 title claims abstract description 99
- 239000004793 Polystyrene Substances 0.000 title claims abstract description 79
- 229920002223 polystyrene Polymers 0.000 title claims abstract description 22
- 238000000926 separation method Methods 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 229920001503 Glucan Polymers 0.000 claims abstract description 50
- 150000002433 hydrophilic molecules Chemical class 0.000 claims abstract description 17
- 238000006243 chemical reaction Methods 0.000 claims description 95
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 75
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 57
- 239000008367 deionised water Substances 0.000 claims description 38
- 229910021641 deionized water Inorganic materials 0.000 claims description 38
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 38
- 238000003756 stirring Methods 0.000 claims description 32
- 238000005303 weighing Methods 0.000 claims description 27
- 238000001914 filtration Methods 0.000 claims description 23
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 14
- 238000004132 cross linking Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 238000000576 coating method Methods 0.000 claims description 11
- 238000001035 drying Methods 0.000 claims description 11
- 230000004048 modification Effects 0.000 claims description 11
- 238000012986 modification Methods 0.000 claims description 11
- 239000011248 coating agent Substances 0.000 claims description 10
- 229920002307 Dextran Polymers 0.000 claims description 9
- 238000004140 cleaning Methods 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- GKIPXFAANLTWBM-UHFFFAOYSA-N epibromohydrin Chemical compound BrCC1CO1 GKIPXFAANLTWBM-UHFFFAOYSA-N 0.000 claims description 8
- 238000004321 preservation Methods 0.000 claims description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 6
- BRLQWZUYTZBJKN-UHFFFAOYSA-N Epichlorohydrin Chemical compound ClCC1CO1 BRLQWZUYTZBJKN-UHFFFAOYSA-N 0.000 claims description 6
- 239000003431 cross linking reagent Substances 0.000 claims description 6
- 239000003960 organic solvent Substances 0.000 claims description 6
- UWFRVQVNYNPBEF-UHFFFAOYSA-N 1-(2,4-dimethylphenyl)propan-1-one Chemical compound CCC(=O)C1=CC=C(C)C=C1C UWFRVQVNYNPBEF-UHFFFAOYSA-N 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 4
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 3
- SEFYJVFBMNOLBK-UHFFFAOYSA-N 2-[2-[2-(oxiran-2-ylmethoxy)ethoxy]ethoxymethyl]oxirane Chemical compound C1OC1COCCOCCOCC1CO1 SEFYJVFBMNOLBK-UHFFFAOYSA-N 0.000 claims description 3
- UMILHIMHKXVDGH-UHFFFAOYSA-N Triethylene glycol diglycidyl ether Chemical compound C1OC1COCCOCCOCCOCC1CO1 UMILHIMHKXVDGH-UHFFFAOYSA-N 0.000 claims description 3
- 229960005237 etoglucid Drugs 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 2
- 235000019441 ethanol Nutrition 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- -1 epibromopropane Chemical compound 0.000 claims 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 abstract description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract description 4
- 238000001212 derivatisation Methods 0.000 abstract description 2
- 239000002861 polymer material Substances 0.000 abstract description 2
- 239000007787 solid Substances 0.000 description 21
- 238000001556 precipitation Methods 0.000 description 14
- 150000001875 compounds Chemical class 0.000 description 12
- 230000002209 hydrophobic effect Effects 0.000 description 9
- 238000000643 oven drying Methods 0.000 description 7
- 238000001179 sorption measurement Methods 0.000 description 7
- LRWZZZWJMFNZIK-UHFFFAOYSA-N 2-chloro-3-methyloxirane Chemical compound CC1OC1Cl LRWZZZWJMFNZIK-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 3
- 239000000725 suspension Chemical group 0.000 description 3
- 230000001588 bifunctional effect Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- CHRJZRDFSQHIFI-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;styrene Chemical compound C=CC1=CC=CC=C1.C=CC1=CC=CC=C1C=C CHRJZRDFSQHIFI-UHFFFAOYSA-N 0.000 description 1
- NYTKHVSYWKBBTE-UHFFFAOYSA-N [Br].C(C)(=O)Cl Chemical compound [Br].C(C)(=O)Cl NYTKHVSYWKBBTE-UHFFFAOYSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- KCXMKQUNVWSEMD-UHFFFAOYSA-N benzyl chloride Chemical compound ClCC1=CC=CC=C1 KCXMKQUNVWSEMD-UHFFFAOYSA-N 0.000 description 1
- 229940073608 benzyl chloride Drugs 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 229920000891 common polymer Polymers 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229920001477 hydrophilic polymer Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000004804 winding Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/12—Powdering or granulating
- C08J3/126—Polymer particles coated by polymer, e.g. core shell structures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2325/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an aromatic carbocyclic ring; Derivatives of such polymers
- C08J2325/02—Homopolymers or copolymers of hydrocarbons
- C08J2325/04—Homopolymers or copolymers of styrene
- C08J2325/08—Copolymers of styrene
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/02—Dextran; Derivatives thereof
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- Chemical Kinetics & Catalysis (AREA)
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- Crystallography & Structural Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
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- Processes Of Treating Macromolecular Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention relates to the technical field of high polymer materials, and provides a preparation method of polystyrene microspheres for hydrophilic biological separation. The modified hydrophilic compound of the linear glucan is coated on the surface of the PS/DVB microsphere to cover a benzene ring and a suspended double bond on the surface of the PS/DVB microsphere, so that the hydrophobicity of the surface of the PS/DVB microsphere is reduced, and the coated hydrophilic compound of the linear glucan introduces a large amount of hydroxyl groups on the surface of the PS/DVB microsphere, is convenient for further derivatization, and can be widely applied to the field of biological separation.
Description
Technical Field
The invention relates to the technical field of high polymer materials, in particular to a preparation method of polystyrene microspheres for hydrophilic biological separation.
Background
Poly (styrene-divinylbenzene) microspheres (PS/DVB microspheres) have wide application in the field of high performance liquid chromatography due to the advantages of good physical and chemical stability, high mechanical strength, high specific surface area and the like, but have hydrophobicity due to a large number of benzene rings and suspended double bonds (Spectroscopy letters,36(6),867 and 874(2000)) on the surfaces of the PS/DVB microspheres, so that non-specific adsorption can be generated on biological macromolecules such as proteins and the like, the application of the biological macromolecules in the biological field is limited, and in order to overcome the defect, hydrophilic modification needs to be carried out on the surfaces of the PS/DVB microspheres to eliminate hydrophobic areas on the surfaces of the PS/DVB microspheres.
At present, the commonly used method for hydrophilic modification of the surface of the PS/DVB microsphere comprises 2 methods: physical adsorption and chemical bonding. Physical adsorption, i.e., the "coating" of one or more layers of "amphiphilic" polymers containing both hydrophobic/hydrophilic regions onto the surface of PS/DVB microspheres followed by post-crosslinking to give stable hydrophilic microspheres (Langmuir,2001,17, 4386-4391; Covalently bound coatings, U.S. Pat. No. 5,503,933 (1996)); chemical bonding usually requires bonding an active group such as benzyl chloride or acetyl chloride (bromine) on the surface of the PS/DVB microsphere, and then reacting with a hydrophilic polymer by utilizing the activity of the group to achieve the purpose of eliminating hydrophobicity. Common polymers for bonding are PVA, PEG, etc., but after the PVA and PEG are grafted onto the surface of PS/DVB microsphere, the PVA or PEG chain is too long to easily "entangle" on the surface of the PS/DVB microsphere, and during the bio-separation process, if the mobile phase has high salt concentration, the phase collapse is easy to occur (Egbert Muller, Chemical Engineering & Technology,2005,28, 1292-.
Disclosure of Invention
Therefore, in view of the above-mentioned disadvantages of the prior art, the present invention is directed to a method for preparing hydrophilic polystyrene microspheres for bioseparation. The invention provides a preparation method of polystyrene microspheres for hydrophilic bioseparation, which comprises the steps of firstly introducing a hydrophobic structural unit into a linear glucan side chain by a chemical modification method, increasing glucan hydrophobic points, then carrying out pi-pi adsorption on a modified linear glucan compound with the added hydrophobic points and a benzene ring or a suspended double bond on the surface of a PS/DVB microsphere by a physical adsorption method, and folding or winding glucan between the hydrophobic points to form a Loop ring after the adjacent hydrophobic points of the modified linear glucan compound are adsorbed on the PS/DVB microsphere, wherein as shown in figure 1, the degree of freedom of hydroxyl groups of the hydrophilic PS/DVB microsphere is increased; the modified linear glucan compound is coated on the surface of the PS/DVB microsphere, and a proper bifunctional group compound is selected and then a hydrophilic layer is crosslinked, so that a large number of hydroxyl groups are introduced to the surface of the PS/DVB microsphere, the aim of reducing the hydrophobicity of the PS/DVB microsphere is fulfilled, and the large number of hydroxyl groups on the surface are convenient for further derivatization reaction.
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of the hydrophilic compound of linear dextran:
weighing a certain amount of linear glucan, adding the linear glucan into deionized water, dissolving, cooling to 40-60 ℃, adding a NaOH solution or KOH solution with a certain concentration, and keeping the concentration of NaOH or KOH in the system to be 1M; adding a certain amount of epichlorohydrin or epibromohydrin, controlling the temperature to be 40-60 ℃, reacting for 2-10h, precipitating the obtained reaction solution in absolute ethyl alcohol or methanol for 2 times, dissolving with deionized water, repeatedly precipitating for 2 times, and finally drying to obtain a modified hydrophilic compound;
(2) hydrophilic coating of PS/DVB microsphere surface:
weighing a certain amount of modified linear glucan hydrophilic compound, adding deionized water for dissolving, adding an organic solvent M1 in a proper proportion, and preparing a solution with a certain concentration for later use; weighing a certain amount of dry PS/DVB microspheres in a reaction bottle, adding the prepared chemically modified linear glucan solution with a certain concentration into the reaction, and stirring; the system is subjected to heat preservation reaction for 6-12h at the temperature of 30-60 ℃; after the reaction is finished, filtering the microspheres by using a sand core funnel, and draining for later use;
(3) crosslinking and cleaning
Transferring the microspheres adsorbing the linear glucan hydrophilic chemicals obtained in the step (2) into a reaction bottle, adding a certain amount of NaOH or KOH solution, and uniformly stirring; continuously adding a certain amount of cross-linking agent M2, heating the system to 40-80 ℃, and reacting for 4-12h under heat preservation; and after the reaction is finished, carrying out suction filtration on the sand core, respectively washing the sand core with methanol or ethanol and a large amount of deionized water, and drying to obtain the hydrophilized PS/DVB microspheres.
The further improvement is that: the organic solvent M1 is isopropanol, dioxane or dimethyl sulfoxide.
The further improvement is that: the crosslinking agent M2 is epichlorohydrin, epoxy bromopropane, ethylene glycol diglycidyl ether, diethylene glycol diglycidyl ether, or triethylene glycol diglycidyl ether.
The further improvement is that: the molecular weight of the linear glucan is 1000-500000, wherein 1000-25000 is preferred.
The further improvement is that: the molar ratio of the epichlorohydrin or the epibromohydrin to the linear glucan is 1-20: 1.
the further improvement is that: the mass ratio of the organic solvent M1 to the deionized water is 1-5: 5.
The further improvement is that: the mass ratio of the modified linear glucan hydrophilic compound to the PS/DVB microspheres is 0.2-2: 1.
The further improvement is that: the mass ratio of the crosslinking agent M2 to the PS/DVB microspheres is 0.2-1: 1.
The invention has the characteristics that:
the polystyrene microsphere for hydrophilic biological separation is characterized in that a benzene ring and a suspension double bond hydrophobic region on the surface of a PS/DVB microsphere are adsorbed with a hydrophobic region in chemically modified linear glucan to form pi-pi adsorption, the linear glucan is adsorbed with the surface of the PS/DVB microsphere, and then a linear glucan compound is fixed on the surface of the PS/DVB microsphere through a post-crosslinking reaction of a bifunctional group to cover the benzene ring and the suspension double bond on the surface of the PS/DVB microsphere, so that the hydrophobicity caused by the residual benzene ring and the suspension double bond on the surface of the PS/DVB microsphere is eliminated; and secondly, the linear glucan forms a compact hydrophilic layer on the surface of the PS/DVB microsphere through post-crosslinking reaction, so that the influence of phase collapse on the separation effect caused by high salt in the field of biological separation by the hydrophilic sphere is avoided. The method also has the following advantages: the synthetic method has the advantages of simplicity, easy control, convenience for amplification production and the like, and has potential application value in the fields of bioseparation, biomedical detection and the like.
Drawings
FIG. 1 is a schematic diagram of adsorption of modified linear glucan on the surface of PS/DVB microspheres
Detailed Description
The following detailed description will be provided for the embodiments of the present invention with reference to specific embodiments, so that how to apply the technical means to solve the technical problems and achieve the technical effects can be fully understood and implemented.
Unless otherwise indicated, the techniques employed in the examples are conventional and well known to those skilled in the art, and the reagents and products employed are also commercially available. The source, trade name and if necessary the constituents of the reagents used are indicated at the first appearance.
Example one
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 5g of linear glucan (with the molecular weight of 10000) into a reaction bottle, adding 40g of deionized water, heating to 50 ℃, stirring until the glucan is completely dissolved, adding NaOH, and keeping the concentration of NaOH in the system to be 1M; slowly dripping 0.05g of epoxy chloropropane, and keeping the temperature of the system at 40 ℃ for reacting for 6 h; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 40g of deionized water for dissolving, adding 10g of isopropanol, uniformly stirring, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reaction for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) to a reaction bottle, adding 20mL of NaOH (3M) solution, and uniformly stirring; continuously adding 1g of epoxy chloropropane into a reaction bottle, and keeping the temperature at 40 ℃ for reaction for 8 hours; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
Example two
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 10g of linear glucan (with molecular weight of 2000) into a reaction bottle, adding 40g of deionized water, heating to 40 ℃, stirring until the glucan is completely dissolved, adding NaOH, and keeping the concentration of NaOH in the system at 1M; slowly dripping 2.2g of epoxy chloropropane, and keeping the temperature of the system at 40 ℃ for reaction for 10 h; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 45g of deionized water for dissolving, adding 5g of isopropanol, stirring uniformly, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reaction for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) to a reaction bottle, adding 20mL of KOH (3M) solution, and uniformly stirring; continuously adding 3g of epoxy bromopropane into the reaction bottle, and reacting for 6 hours at the temperature of 40 ℃; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
EXAMPLE III
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 8g of linear glucan (with the molecular weight of 1500) into a reaction bottle, adding 40g of deionized water, heating to 50 ℃, stirring until the glucan is completely dissolved, adding KOH, and keeping the concentration of the KOH in the system to be 1M; slowly dripping 5.8g of epoxy bromopropane, and reacting for 6 hours at the temperature of 50 ℃; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 30g of deionized water for dissolving, adding 10g of isopropanol, uniformly stirring, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reaction for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) to a reaction bottle, adding 20mL of KOH (3M) solution, and uniformly stirring; continuously adding 3g of ethylene glycol diglycidyl ether into a reaction bottle, and carrying out heat preservation reaction at 40 ℃ for 6 h; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
Example four
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 5g of linear glucan (with molecular weight of 6000) in a reaction bottle, adding 40g of deionized water, heating to 40 ℃, stirring until the glucan is completely dissolved, adding NaOH, and keeping the concentration of NaOH in the system at 1M; slowly dripping 0.46g of epoxy chloropropane, and keeping the temperature of the system at 40 ℃ for reacting for 8 h; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 40g of deionized water for dissolving, adding 10g of dioxane, stirring uniformly, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reacting for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) into a reaction bottle, adding 20mL of KOH (3M) solution, and uniformly stirring; continuously adding 5g of diethylene glycol diglycidyl ether into the reaction flask, and carrying out heat preservation reaction at 40 ℃ for 6 h; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
EXAMPLE five
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 5g of linear glucan (with the molecular weight of 25000) into a reaction bottle, adding 40g of deionized water, heating to 40 ℃, stirring until the glucan is completely dissolved, adding NaOH, and keeping the concentration of NaOH in a system to be 1M; slowly dripping 0.13g of epoxy chloropropane, and keeping the temperature of the system at 40 ℃ for reacting for 6 h; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 40g of deionized water for dissolving, adding 5g of isopropanol, uniformly stirring, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reaction for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) to a reaction bottle, adding 20mL of KOH (3M) solution, and uniformly stirring; continuously adding 2g of triethylene glycol diglycidyl ether into a reaction bottle, and carrying out heat preservation reaction at 40 ℃ for 6 h; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
EXAMPLE six
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 1.2g of linear glucan (molecular weight of 60000) in a reaction bottle, adding 20g of deionized water, heating to 50 ℃, stirring until the glucan is completely dissolved, adding NaOH, and keeping the concentration of NaOH in the system at 1M; slowly dripping 0.05g of epoxy chloropropane, and keeping the temperature of the system at 50 ℃ for reaction for 4 h; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 40g of deionized water for dissolving, adding 5g of dimethyl sulfoxide, uniformly stirring, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reaction for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) to a reaction bottle, adding 20mL of KOH (3M) solution, and uniformly stirring; continuously adding 3g of ethylene glycol diglycidyl ether into a reaction bottle, and carrying out heat preservation reaction at 40 ℃ for 6 h; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
EXAMPLE seven
A preparation method of polystyrene microspheres for hydrophilic biological separation comprises the following steps:
(1) modification of hydrophilic compounds of linear dextrans
Weighing 0.5g of linear glucan (with the molecular weight of 250000) into a reaction bottle, adding 20g of deionized water, heating to 60 ℃, stirring until the glucan is completely dissolved, adding NaOH, and keeping the concentration of NaOH in a system to be 1M; slowly dripping 0.05g of epoxy bromopropane, and keeping the temperature of the system at 60 ℃ for reaction for 2 hours; after the reaction is finished, pouring the reaction solution into methanol for precipitation, and filtering out solids; dissolving the solid in deionized water, adding into methanol for precipitation, filtering to obtain solid, and oven drying.
(2) Hydrophilic coating of PS/DVB microsphere surface
Weighing 5g of dry PS/DVB microspheres and a reaction bottle; weighing the modified linear glucan compound obtained in the step (1) and a beaker, adding 40g of deionized water for dissolving, adding 5g of isopropanol, uniformly stirring, transferring the solution into a reaction bottle, keeping the temperature of the system at 40 ℃, and stirring for reaction for 10 hours; after the reaction is finished, filtering out microspheres, and draining for later use.
(3) Crosslinking and cleaning
Transferring the microspheres obtained in the step (2) to a reaction bottle, adding 20mL of KOH (3M) solution, and uniformly stirring; 2.5g of ethylene glycol diglycidyl ether is continuously added into a reaction bottle, and the reaction is carried out for 6 hours at the temperature of 40 ℃; after the reaction is finished, washing with methanol and deionized water respectively, and drying to obtain the hydrophilized PS/DVB microspheres.
Claims (8)
1. A preparation method of polystyrene microspheres for hydrophilic biological separation is characterized by comprising the following steps: the method comprises the following steps:
(1) modification of the hydrophilic compound of linear dextran:
weighing a certain amount of linear glucan, adding the linear glucan into deionized water, dissolving, cooling to 40-60 ℃, adding a NaOH solution or KOH solution with a certain concentration, and keeping the concentration of NaOH or KOH in the system at 1M; adding a certain amount of epichlorohydrin or epibromohydrin, controlling the temperature to be 40-60 ℃, reacting for 2-10h, precipitating the obtained reaction solution in absolute ethyl alcohol or methanol for 2 times, dissolving with deionized water, repeatedly precipitating for 2 times, and finally drying to obtain a modified hydrophilic compound;
(2) hydrophilic coating of PS/DVB microsphere surface:
weighing a certain amount of modified linear glucan hydrophilic compound, adding deionized water for dissolving, adding an organic solvent M1 in a proper proportion, and preparing a solution with a certain concentration for later use; weighing a certain amount of dry PS/DVB microspheres in a reaction bottle, adding the prepared chemically modified linear glucan solution with a certain concentration into the reaction, and stirring; the system is subjected to heat preservation reaction for 6-12h at the temperature of 30-60 ℃; after the reaction is finished, filtering the microspheres by using a sand core funnel, and draining for later use;
(3) crosslinking and cleaning
Transferring the microspheres adsorbing the linear glucan hydrophilic chemicals obtained in the step (2) into a reaction bottle, adding a certain amount of NaOH or KOH solution, and uniformly stirring; continuously adding a certain amount of cross-linking agent M2, heating the system to 40-80 ℃, and reacting for 4-12h under heat preservation; and after the reaction is finished, carrying out suction filtration on the sand core, respectively washing the sand core with methanol or ethanol and a large amount of deionized water, and drying to obtain the hydrophilized PS/DVB microspheres.
2. The method of claim 1, wherein the organic solvent M1 is isopropanol, dioxane, or dimethyl sulfoxide.
3. The method for preparing hydrophilic polystyrene microspheres for biological separation according to claim 1, wherein the crosslinking agent M2 is epichlorohydrin, epibromopropane, ethylene glycol diglycidyl ether, diethylene glycol diglycidyl ether, or triethylene glycol diglycidyl ether.
4. The method for preparing polystyrene microspheres for hydrophilic biological separation according to claim 1, wherein the molecular weight of the linear glucan is 1000-500000, preferably 1000-25000.
5. The preparation method of the hydrophilic polystyrene microsphere for bioseparation according to claim 1, wherein the molar ratio of the epichlorohydrin or the epibromohydrin to the linear glucan is 1-20: 1.
6. the method for preparing polystyrene microspheres for hydrophilic bioseparation according to claim 1, wherein the mass ratio of the organic solvent M1 to deionized water is 1-5: 5.
7. The preparation method of the polystyrene microsphere for the hydrophilic bioseparation, according to claim 1, is characterized in that the mass ratio of the modified linear glucan hydrophilic compound to the PS/DVB microsphere is 0.2-2: 1.
8. The preparation method of the polystyrene microsphere for the hydrophilic bioseparation, according to claim 1, is characterized in that the mass ratio of the cross-linking agent M2 to the PS/DVB microsphere is 0.2-1: 1.
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