CN1844513A - Preparation of acrylon nano fiber membrane carrier materials and use thereof - Google Patents
Preparation of acrylon nano fiber membrane carrier materials and use thereof Download PDFInfo
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- CN1844513A CN1844513A CN 200610049954 CN200610049954A CN1844513A CN 1844513 A CN1844513 A CN 1844513A CN 200610049954 CN200610049954 CN 200610049954 CN 200610049954 A CN200610049954 A CN 200610049954A CN 1844513 A CN1844513 A CN 1844513A
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- fiber membrane
- carrier materials
- reactive group
- nano fiber
- membrane carrier
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- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 title claims description 72
- 239000012528 membrane Substances 0.000 title claims description 63
- 239000012876 carrier material Substances 0.000 title claims description 62
- 239000002121 nanofiber Substances 0.000 title claims description 58
- HGINCPLSRVDWNT-UHFFFAOYSA-N acrylaldehyde Natural products C=CC=O HGINCPLSRVDWNT-UHFFFAOYSA-N 0.000 title claims description 37
- 238000009987 spinning Methods 0.000 claims abstract description 37
- 102000004190 Enzymes Human genes 0.000 claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 claims abstract description 23
- 238000010041 electrostatic spinning Methods 0.000 claims abstract description 21
- 239000000463 material Substances 0.000 claims abstract description 20
- 229920002239 polyacrylonitrile Polymers 0.000 claims abstract description 16
- 239000002904 solvent Substances 0.000 claims abstract description 8
- 239000000126 substance Substances 0.000 claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 claims abstract description 7
- 229920006243 acrylic copolymer Polymers 0.000 claims description 17
- 238000000034 method Methods 0.000 claims description 17
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 16
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 15
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 claims description 12
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 claims description 12
- 239000011976 maleic acid Substances 0.000 claims description 12
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 claims description 12
- 229920001577 copolymer Polymers 0.000 claims description 10
- 102000004357 Transferases Human genes 0.000 claims description 8
- 108090000992 Transferases Proteins 0.000 claims description 8
- -1 antibody Proteins 0.000 claims description 8
- 150000001718 carbodiimides Chemical class 0.000 claims description 8
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 8
- 239000007822 coupling agent Substances 0.000 claims description 7
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 6
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 claims description 6
- 102000004882 Lipase Human genes 0.000 claims description 5
- 108090001060 Lipase Proteins 0.000 claims description 5
- 239000004367 Lipase Substances 0.000 claims description 5
- 235000019421 lipase Nutrition 0.000 claims description 5
- 102000008186 Collagen Human genes 0.000 claims description 4
- 108010035532 Collagen Proteins 0.000 claims description 4
- 108010010803 Gelatin Proteins 0.000 claims description 4
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims description 4
- 102000004877 Insulin Human genes 0.000 claims description 4
- 108090001061 Insulin Proteins 0.000 claims description 4
- 238000010521 absorption reaction Methods 0.000 claims description 4
- 238000004140 cleaning Methods 0.000 claims description 4
- 229920001436 collagen Polymers 0.000 claims description 4
- 229920000159 gelatin Polymers 0.000 claims description 4
- 239000008273 gelatin Substances 0.000 claims description 4
- 235000019322 gelatine Nutrition 0.000 claims description 4
- 235000011852 gelatine desserts Nutrition 0.000 claims description 4
- 229920000669 heparin Polymers 0.000 claims description 4
- 229960002897 heparin Drugs 0.000 claims description 4
- 229940125396 insulin Drugs 0.000 claims description 4
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 claims description 3
- HJBUBXIDMQBSQW-UHFFFAOYSA-N 4-(4-diazoniophenyl)benzenediazonium Chemical compound C1=CC([N+]#N)=CC=C1C1=CC=C([N+]#N)C=C1 HJBUBXIDMQBSQW-UHFFFAOYSA-N 0.000 claims description 3
- XZMCDFZZKTWFGF-UHFFFAOYSA-N Cyanamide Chemical compound NC#N XZMCDFZZKTWFGF-UHFFFAOYSA-N 0.000 claims description 3
- BRLQWZUYTZBJKN-UHFFFAOYSA-N Epichlorohydrin Chemical compound ClCC1CO1 BRLQWZUYTZBJKN-UHFFFAOYSA-N 0.000 claims description 3
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 3
- YACXKLPYAFZMHA-UHFFFAOYSA-N [Cl].C1(=CC=C(C=C1)S(=O)(=O)O)C Chemical compound [Cl].C1(=CC=C(C=C1)S(=O)(=O)O)C YACXKLPYAFZMHA-UHFFFAOYSA-N 0.000 claims description 3
- 229910021529 ammonia Inorganic materials 0.000 claims description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 3
- VBNJLNQSWSLPEP-UHFFFAOYSA-N aniline 1-ethylsulfonylethane Chemical compound CCS(=O)(=O)CC.NC1=CC=CC=C1 VBNJLNQSWSLPEP-UHFFFAOYSA-N 0.000 claims description 3
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 claims description 3
- 150000002148 esters Chemical class 0.000 claims description 3
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 2
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 claims description 2
- 102000057234 Acyl transferases Human genes 0.000 claims description 2
- 108700016155 Acyl transferases Proteins 0.000 claims description 2
- 229920001817 Agar Polymers 0.000 claims description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 2
- 229920001287 Chondroitin sulfate Polymers 0.000 claims description 2
- 102000000634 Cytochrome c oxidase subunit IV Human genes 0.000 claims description 2
- 108050008072 Cytochrome c oxidase subunit IV Proteins 0.000 claims description 2
- 101710088194 Dehydrogenase Proteins 0.000 claims description 2
- 108010073385 Fibrin Proteins 0.000 claims description 2
- 102000009123 Fibrin Human genes 0.000 claims description 2
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 claims description 2
- 108010022355 Fibroins Proteins 0.000 claims description 2
- 241000206672 Gelidium Species 0.000 claims description 2
- 229920001503 Glucan Polymers 0.000 claims description 2
- 102000051366 Glycosyltransferases Human genes 0.000 claims description 2
- 108700023372 Glycosyltransferases Proteins 0.000 claims description 2
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 claims description 2
- 102000004020 Oxygenases Human genes 0.000 claims description 2
- 108090000417 Oxygenases Proteins 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims description 2
- 108091005804 Peptidases Proteins 0.000 claims description 2
- 102000003992 Peroxidases Human genes 0.000 claims description 2
- 239000004365 Protease Substances 0.000 claims description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 2
- 102000003929 Transaminases Human genes 0.000 claims description 2
- 108090000340 Transaminases Proteins 0.000 claims description 2
- 229920006322 acrylamide copolymer Polymers 0.000 claims description 2
- 235000010419 agar Nutrition 0.000 claims description 2
- 235000010443 alginic acid Nutrition 0.000 claims description 2
- 229920000615 alginic acid Polymers 0.000 claims description 2
- 229940024606 amino acid Drugs 0.000 claims description 2
- 239000000427 antigen Substances 0.000 claims description 2
- 102000036639 antigens Human genes 0.000 claims description 2
- 108091007433 antigens Proteins 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 235000010418 carrageenan Nutrition 0.000 claims description 2
- 239000000679 carrageenan Substances 0.000 claims description 2
- 229920001525 carrageenan Polymers 0.000 claims description 2
- 229940113118 carrageenan Drugs 0.000 claims description 2
- 229940059329 chondroitin sulfate Drugs 0.000 claims description 2
- 229950003499 fibrin Drugs 0.000 claims description 2
- 229920002674 hyaluronan Polymers 0.000 claims description 2
- 229960003160 hyaluronic acid Drugs 0.000 claims description 2
- 229920003145 methacrylic acid copolymer Polymers 0.000 claims description 2
- 229940117841 methacrylic acid copolymer Drugs 0.000 claims description 2
- 150000007523 nucleic acids Chemical class 0.000 claims description 2
- 102000039446 nucleic acids Human genes 0.000 claims description 2
- 108020004707 nucleic acids Proteins 0.000 claims description 2
- 230000001590 oxidative effect Effects 0.000 claims description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 2
- 239000010452 phosphate Substances 0.000 claims description 2
- 229910052717 sulfur Inorganic materials 0.000 claims description 2
- 239000011593 sulfur Substances 0.000 claims description 2
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 claims description 2
- 239000000835 fiber Substances 0.000 abstract description 14
- 229920002972 Acrylic fiber Polymers 0.000 abstract 2
- 238000005070 sampling Methods 0.000 abstract 1
- 238000005204 segregation Methods 0.000 abstract 1
- 238000001179 sorption measurement Methods 0.000 abstract 1
- 208000012886 Vertigo Diseases 0.000 description 32
- 239000000243 solution Substances 0.000 description 25
- 239000008367 deionised water Substances 0.000 description 6
- 239000008055 phosphate buffer solution Substances 0.000 description 5
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- 239000004952 Polyamide Substances 0.000 description 1
- 229940061720 alpha hydroxy acid Drugs 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
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- 238000001816 cooling Methods 0.000 description 1
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- 239000002657 fibrous material Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229910052615 phyllosilicate Inorganic materials 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920000570 polyether Polymers 0.000 description 1
- 229920005594 polymer fiber Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
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Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
This invention relates to the preparation method for the acrylic fibers nanometer fiber film with reaction group, which includes following steps: dissolving the polyacrylonitrile copolymer with reaction group in the solvent to make the solvent become to the transparent solution, then adding the solution in the micro scale sampling device, connecting the spinning head with the high-voltage power, wherein the voltage is 5 kv-20 kv, electrostatic spinning to obtain the acrylic fibers nanometer fiber film materials with reaction group. This invention fixes the biological molecule or enzyme on the material surface by physics adsorption or chemical covalent binding connection. The material can be used in organizing project frame, the enzyme's fixing, the biological molecule's segregation.
Description
Technical field
The invention belongs to the technology of preparing of the acrylon nano fiber membrane carrier materials that contains reactive group and be used for biomolecule and method that enzyme is fixing.
Background technology
Electrostatic spinning technique is spinning under electric field with Polymer Solution or melt, owing to electrostatic interaction produces stretching, obtains superfine fibre through cooling or solvent evaporates.In the electrostatic spinning process, can form the superfine nano tunica fibrosa that superfine fibre can be piled up.
Existing electrostatic spinning membrane preparation method has following several:
CN 200410019230.9; 200410019231.3 shitosan, poly-Alpha-hydroxy acid esters/shitosan electrostatic spinning are prepared superfine fibre film, and this material can be used for tissue repair.
Macromolecular material electrostatic spinnings such as CN 03131586.0 usefulness polyester, polyamide, polyethers obtain nano fibrous membrane, at the common polymer fiber material of its surface stack, can be used as protective materials.
CA 2494865; 2386674 prepare polymer nanofiber with the polymeric material electrostatic spinning is used for pharmaceutical carrier or tissue renovation material.
US 20050014252; 20040185737; But 20030137083 usefulness Biodegradable material bioabsorbable materials, silk class material, conducting polymer electrostatic spinning obtain superfine fibre film.
US 20040013873; 20020122840 by electrostatic spinning, the polymer fiber film of preparation porous.
On the other hand, various biomolecule are fixed on the carrier material, important use are all arranged in fields such as bio-molecular separation, enzymic catalytic reaction, organizational projects.
The preparation of carrier material has following several:
CN01126606.6; CN200410025793.9 makes carrier material with pearl shitosan, phyllosilicate, is used for the immobilization of enzyme.
In above-mentioned prior art, do not find effectively to utilize the electrostatic spinning technique preparation to be used for biomolecule and the fixing carrier material of enzyme as yet.The specific area of current material is less, is difficult to increase substantially biomolecule and the fixing quantity of enzyme, is unfavorable for reducing production costs.
Summary of the invention
The purpose of this invention is to provide a kind of utilize electrostatic spinning technique preparation be used for biomolecule and the fixing carrier material of enzyme, have extensive use at aspects such as the immobilization of tissue engineering bracket, enzyme, bio-molecular separation and be worth.
For achieving the above object, the technical scheme of taking: adopt the preparation of electrostatic spinning technique technology to contain the acrylon nano fiber membrane carrier materials of reactive group, be that the acrylonitrile copolymer that will contain reactive group is dissolved in the solvent, become spinning solution, carry out electrostatic spinning, obtain containing the acrylon nano fiber membrane carrier materials of reactive group, fibre diameter is between 30 nanometers-5 micron; Method by chemical covalent bond connection or physical absorption is fixed on material surface with biomolecule or enzyme.The method that is used for biomolecule mobilization: be that the acrylon nano fiber membrane carrier materials of reactive group is put into the solution that contains biomolecule, add coupling agent reaction back and take out this nano fibrous membrane, obtain the acrylon nano fiber membrane carrier materials that contains reactive group of biomolecule mobilization after having cleaned; The method that is used for enzyme immobilization: be that the acrylon nano fiber membrane carrier materials that contains reactive group is put into solution, add coupling agent reaction back and take out the acrylon nano fiber membrane carrier materials that contains reactive group, after the cleaning, putting into the solution that contains enzyme reacts, membrane material is taken out cleaning, promptly get the acrylon nano fiber membrane carrier materials that contains reactive group of enzyme immobilization.
Remarkable advantage of the present invention and effect:
1) contain the acrylon nano fiber membrane carrier materials of reactive group, this kind fiber is superfine, and diameter has very big specific area between 30 nanometers-5 micron, help improving the immobilized quantity of biomolecule and enzyme;
2) to contain the technology of acrylon nano fiber membrane carrier materials of reactive group simple in preparation, and the acrylonitrile copolymer dissolving that will contain reactive group obtains spinning solution, carries out electrostatic spinning, just can obtain containing the acrylon nano fiber membrane carrier materials of reactive group;
3) acrylon nano fiber membrane carrier materials that contains reactive group can be processed into tubulose, bulk, strip, thread, graininess, individual layer is membranaceous, multilayer is membranaceous, is used for the aspect such as immobilization, bio-molecular separation of tissue repair, tissue engineering bracket, enzyme.
The specific embodiment
The method of the preparation of the acrylon nano fiber membrane carrier materials that contains reactive group of the present invention, be that the acrylonitrile copolymer that will contain reactive group is dissolved in and becomes clear solution in the solvent, obtain spinning solution, join in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, voltage is 5 kilovolts-20 kilovolts, carry out electrostatic spinning, obtain containing the acrylon nano fiber membrane carrier materials of reactive group, fibre diameter is between 30 nanometers-5 micron.
The acrylonitrile copolymer that wherein contains reactive group is any one in acrylonitrile and maleic acid, acrylonitrile and acrylic copolymer, acrylonitrile and methacrylic acid copolymer, acrylonitrile and acrylamide copolymer, acrylonitrile and the methacrylamide copolymer; The solvent that the acrylonitrile copolymer dissolving that will contain reactive group obtains spinning solution is any one in dimethyl sulfoxide (DMSO), dimethylformamide, the dimethyl acetyl ammonia; Connect by physical absorption or chemical covalent bond, the biomolecule that is fixed in the acrylon nano fiber membrane carrier materials surface of this kind reactive group is any one in collagen, gelatin, antibody, antigen, nucleic acid, poly-a-amino acid, hyaluronic acid, chondroitin sulfate, carrageenan, alginates, agar-agar, glucan, fibrin, silk-fibroin, insulin, the heparin; The enzyme that is fixed in this kind material surface is lipase, protease, dehydrogenase, oxidizing ferment, peroxidase, oxygenase, cytochrome oxidase, carbon back transferase, ketaldonyl transferase, acyltransferase, glycosyl transferase, contain in amino transferase, phosphate transferase, the sulfur-containing group transferase any one; Connect by chemical covalent bond, the coupling agent that is fixed in the biomolecule of this kind material surface or enzyme is any one in the inferior acyl ammonia of carbodiimide, hydroxyl amber, glutaraldehyde, cyanogen bromide, hydrazine hydrate, p-methyl benzenesulfonic acid chlorine, chloropropylene oxide, thionyl chloride, sulfuric acid ester ethyl sulfone aniline, carbimide, the bis-diazotized benzidine.
Below introduce the embodiment of preparation material of the present invention:
Example 1: contain the preparation of the acrylon nano fiber membrane carrier materials of reactive group: with 6 grammes per square metre average molecular weights is that 100,000 acrylonitrile and maleic acid are dissolved in and become clear solution in the dimethylformamide, obtain spinning solution, join in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, distance between spinning head and the accepted thing is about 15 centimetres, the spinning solution flow velocity is 5ml/h, voltage is 10 kilovolts, carry out electrostatic spinning, obtain acrylonitrile and maleic acid nano fiber membrane carrier materials, fibre diameter is 100-500nm.
The immobilization of biomolecule: with the acrylonitrile of 1 gram and the phosphate buffer solution (pH=7) of the maleic acid nano fiber membrane carrier materials 30mg/ml collagen of putting into 100 milliliters, add carbodiimide 1 gram, react and take out acrylonitrile and maleic acid nano fiber membrane carrier materials after 24 hours, use washed with de-ionized water, obtain immobilized acrylonitrile of collagen and maleic acid nano fiber membrane carrier materials.
Example 2: contain the preparation of the acrylon nano fiber membrane carrier materials of reactive group: with 5 grammes per square metre average molecular weights is that 150,000 acrylonitrile and acrylic copolymer are dissolved in and become clear solution in the dimethyl sulfoxide (DMSO), obtain spinning solution, join in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, distance between spinning head and the accepted thing is about 20 centimetres, the spinning solution flow velocity is 10ml/h, voltage is 12 kilovolts, carry out electrostatic spinning, obtain acrylonitrile and acrylic copolymer nano fiber membrane carrier materials, fibre diameter is 150-600nm.
The immobilization of biomolecule: with the acrylonitrile of 1 gram and the phosphate buffer solution (pH=7) of the acrylic copolymer nano fiber membrane carrier materials 35mg/ml gelatin of putting into 100 milliliters, add carbodiimide 1 gram, react and take out acrylonitrile and acrylic copolymer nano fiber membrane carrier materials after 24 hours, use washed with de-ionized water, obtain immobilized acrylonitrile of gelatin and acrylic copolymer nano fiber membrane carrier materials.
Example 3: contain the preparation of the acrylon nano fiber membrane carrier materials of reactive group: with 8 grammes per square metre average molecular weights is that 80,000 acrylonitrile and maleic acid are dissolved in and become clear solution in the dimethyl sulfoxide (DMSO), obtain spinning solution, join in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, distance between spinning head and the accepted thing is about 16 centimetres, the spinning solution flow velocity is 6ml/h, voltage is 12 kilovolts, carry out electrostatic spinning, obtain acrylonitrile and maleic acid nano fiber membrane carrier materials, fibre diameter is 120-500nm.
The immobilization of biomolecule: with the acrylonitrile of 1 gram and the phosphate buffer solution (pH=7) of the maleic acid nano fiber membrane carrier materials 20mg/ml heparin of putting into 100 milliliters, add carbodiimide 1 gram, react and take out acrylonitrile and maleic acid nano fiber membrane carrier materials after 24 hours, use washed with de-ionized water, obtain immobilized acrylonitrile of heparin and maleic acid nano fiber membrane carrier materials.
Example 4: contain the preparation of the acrylon nano fiber membrane carrier materials of reactive group: with 6 grammes per square metre average molecular weights is that 150,000 acrylonitrile and acrylic copolymer are dissolved in and become clear solution in the dimethylformamide, obtain spinning solution, join in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, distance between spinning head and the accepted thing is about 18 centimetres, the spinning solution flow velocity is 5ml/h, voltage is 15 kilovolts, carry out electrostatic spinning, obtain acrylonitrile and acrylic copolymer nano fiber membrane carrier materials, fibre diameter is 160-600nm.
The immobilization of enzyme: the acrylonitrile and the acrylic copolymer nano fiber membrane carrier materials of 1 gram are put into 100 milliliters of phosphate buffer solutions (pH=7), add carbodiimide 1 gram, react and take out acrylonitrile and acrylic copolymer nano fiber membrane carrier materials after 24 hours, use washed with de-ionized water, the phosphate buffer solution (pH=7) of putting into 100 milliliters 30mg/ml lipase reacted 6 hours, use washed with de-ionized water, obtain the acrylonitrile and the acrylic copolymer nano fiber membrane carrier materials of lipase immobilization, the activity of lipase is left 25-40%.
Example 5
Contain the preparation of the acrylon nano fiber membrane carrier materials of reactive group: with 5 grammes per square metre average molecular weights is that 150,000 acrylonitrile and acrylic copolymer are dissolved in and become clear solution in the dimethylformamide, obtain spinning solution, join in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, distance between spinning head and the accepted thing is about 15 centimetres, the spinning solution flow velocity is 5ml/h, voltage is 10 kilovolts, carry out electrostatic spinning, obtain acrylonitrile and acrylic copolymer nano fiber membrane carrier materials, fibre diameter is 100-600nm.
The immobilization of biomolecule: with the acrylonitrile of 1 gram and the phosphate buffer solution (pH=7) of the acrylic copolymer nano fiber membrane carrier materials 30mg/ml insulin of putting into 100 milliliters, add carbodiimide 1 gram, react and take out acrylonitrile and acrylic copolymer nano fiber membrane carrier materials after 24 hours, use washed with de-ionized water, obtain immobilized acrylonitrile of insulin and acrylic copolymer nano fiber membrane carrier materials.
Claims (9)
1, a kind of method of preparation of the acrylon nano fiber membrane carrier materials that contains reactive group, the acrylonitrile copolymer that will contain reactive group is dissolved in becomes clear solution in the solvent, obtain spinning solution, add in the microsyringe, be connected to spinning head, spinning head is connected with high voltage source, voltage is 5 kilovolts-20 kilovolts, carry out electrostatic spinning, obtain containing the acrylon nano fiber membrane carrier materials of reactive group.
2, a kind of acrylon nano fiber membrane carrier materials that contains reactive group according to claim 1 is characterized in that: the acrylonitrile copolymer that contains reactive group is any one in acrylonitrile and maleic acid, acrylonitrile and acrylic copolymer, acrylonitrile and methacrylic acid copolymer, acrylonitrile and acrylamide copolymer, acrylonitrile and the methacrylamide copolymer.
3, a kind of preparation method who contains the acrylon nano fiber membrane carrier materials of reactive group according to claim 1, it is characterized in that will contain the acrylonitrile copolymer dissolving of the reactive group solvent that obtains spinning solution be in dimethyl sulfoxide (DMSO), dimethylformamide, the dimethyl acetyl ammonia any one.
4, a kind of acrylon nano fiber membrane carrier materials that contains reactive group is used for the fixing method of biomolecule, it is characterized in that: be that the acrylon nano fiber membrane carrier materials that contains reactive group is put into the solution that contains biomolecule, add coupling agent reaction back and take out this nano fibrous membrane, obtain the acrylon nano fiber membrane carrier materials of the reactive group of biomolecule mobilization after having cleaned.
5, a kind of acrylon nano fiber membrane carrier materials that contains reactive group according to claim 4 is used for the fixing method of biomolecule, it is characterized in that: connect by physical absorption or chemical covalent bond, the biomolecule that is fixed in this kind material surface is any one in collagen, gelatin, antibody, antigen, nucleic acid, poly-a-amino acid, hyaluronic acid, chondroitin sulfate, carrageenan, alginates, agar-agar, glucan, fibrin, silk-fibroin, insulin, the heparin.
6, a kind of acrylon nano fiber membrane carrier materials that contains reactive group is used for the fixing method of enzyme, it is characterized in that: be that the acrylon nano fiber membrane carrier materials of reactive group is put into the solution that contains coupling agent, the acrylon nano fiber membrane carrier materials that contains reactive group is taken out in the reaction back, after the cleaning, putting into the solution that contains enzyme reacts, membrane material is taken out cleaning, promptly get the acrylon nano fiber membrane carrier materials of the reactive group of enzyme immobilization.
7, a kind of acrylon nano fiber membrane carrier materials that contains reactive group according to claim 6 is used for the fixing method of enzyme; it is characterized in that: connect by physical absorption or chemical covalent bond, the enzyme that is fixed in this kind material surface is lipase, protease, dehydrogenase, oxidizing ferment, peroxidase, oxygenase, cytochrome oxidase, carbon back transferase, ketaldonyl transferase, acyltransferase, glycosyl transferase, contain in amino transferase, phosphate transferase, the sulfur-containing group transferase any one.
8, a kind of acrylon nano fiber membrane carrier materials that contains reactive group according to claim 4 is used for the fixing method of biomolecule, it is characterized in that: connect by chemical covalent bond, the coupling agent that is used for fixing biomolecule is any one in the inferior acyl ammonia of carbodiimide, hydroxyl amber, glutaraldehyde, cyanogen bromide, hydrazine hydrate, p-methyl benzenesulfonic acid chlorine, chloropropylene oxide, thionyl chloride, sulfuric acid ester ethyl sulfone aniline, carbimide, the bis-diazotized benzidine.
9, a kind of acrylon nano fiber membrane carrier materials that contains reactive group according to claim 6 is used for the fixing method of enzyme, it is characterized in that: connect by chemical covalent bond, the coupling agent that is used for fixing biomolecule is any one in the inferior acyl ammonia of carbodiimide, hydroxyl amber, glutaraldehyde, cyanogen bromide, hydrazine hydrate, p-methyl benzenesulfonic acid chlorine, chloropropylene oxide, thionyl chloride, sulfuric acid ester ethyl sulfone aniline, carbimide, the bis-diazotized benzidine.
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