CN101671395A - Method for quickly immobilizing proteins or amino-contained molecules on aminated surface - Google Patents
Method for quickly immobilizing proteins or amino-contained molecules on aminated surface Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 38
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 29
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 29
- 230000003100 immobilizing effect Effects 0.000 title claims abstract description 18
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 80
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 16
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229910052710 silicon Inorganic materials 0.000 claims abstract description 13
- 239000010703 silicon Substances 0.000 claims abstract description 13
- HOGDNTQCSIKEEV-UHFFFAOYSA-N n'-hydroxybutanediamide Chemical compound NC(=O)CCC(=O)NO HOGDNTQCSIKEEV-UHFFFAOYSA-N 0.000 claims abstract description 12
- -1 poly-carboxyl compound Chemical class 0.000 claims abstract description 12
- 239000007790 solid phase Substances 0.000 claims abstract description 8
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000010931 gold Substances 0.000 claims abstract description 4
- 229910052737 gold Inorganic materials 0.000 claims abstract description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 28
- 238000007306 functionalization reaction Methods 0.000 claims description 19
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 12
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 7
- 238000006557 surface reaction Methods 0.000 claims description 7
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 6
- JFCQEDHGNNZCLN-UHFFFAOYSA-N glutaric acid Chemical compound OC(=O)CCCC(O)=O JFCQEDHGNNZCLN-UHFFFAOYSA-N 0.000 claims description 6
- 229920001661 Chitosan Polymers 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 239000011521 glass Substances 0.000 claims description 4
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 claims description 3
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 claims description 3
- HCPOCMMGKBZWSJ-UHFFFAOYSA-N ethyl 3-hydrazinyl-3-oxopropanoate Chemical compound CCOC(=O)CC(=O)NN HCPOCMMGKBZWSJ-UHFFFAOYSA-N 0.000 claims description 3
- 235000006408 oxalic acid Nutrition 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims description 3
- 229940095064 tartrate Drugs 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
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 claims description 2
- JDXQWYKOKYUQDN-UHFFFAOYSA-N 3-hydroxypyrrolidine-2,5-dione Chemical compound OC1CC(=O)NC1=O JDXQWYKOKYUQDN-UHFFFAOYSA-N 0.000 claims description 2
- XZMCDFZZKTWFGF-UHFFFAOYSA-N Cyanamide Chemical compound NC#N XZMCDFZZKTWFGF-UHFFFAOYSA-N 0.000 claims description 2
- KGWDUNBJIMUFAP-KVVVOXFISA-N Ethanolamine Oleate Chemical compound NCCO.CCCCCCCC\C=C/CCCCCCCC(O)=O KGWDUNBJIMUFAP-KVVVOXFISA-N 0.000 claims description 2
- NSOXQYCFHDMMGV-UHFFFAOYSA-N Tetrakis(2-hydroxypropyl)ethylenediamine Chemical compound CC(O)CN(CC(C)O)CCN(CC(C)O)CC(C)O NSOXQYCFHDMMGV-UHFFFAOYSA-N 0.000 claims description 2
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 claims description 2
- 229960002442 glucosamine Drugs 0.000 claims description 2
- 229920002674 hyaluronan Polymers 0.000 claims description 2
- 229960003160 hyaluronic acid Drugs 0.000 claims description 2
- 229920002401 polyacrylamide Polymers 0.000 claims description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 2
- 150000001718 carbodiimides Chemical class 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 102000008102 Ankyrins Human genes 0.000 description 11
- 108010049777 Ankyrins Proteins 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- 238000005576 amination reaction Methods 0.000 description 7
- 150000001299 aldehydes Chemical class 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 230000002939 deleterious effect Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000003912 environmental pollution Methods 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
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- 239000007787 solid Substances 0.000 description 3
- CNODSORTHKVDEM-UHFFFAOYSA-N 4-trimethoxysilylaniline Chemical compound CO[Si](OC)(OC)C1=CC=C(N)C=C1 CNODSORTHKVDEM-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
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- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 2
- RINCXYDBBGOEEQ-UHFFFAOYSA-N succinic anhydride Chemical compound O=C1CCC(=O)O1 RINCXYDBBGOEEQ-UHFFFAOYSA-N 0.000 description 2
- KUCOHFSKRZZVRO-UHFFFAOYSA-N terephthalaldehyde Chemical compound O=CC1=CC=C(C=O)C=C1 KUCOHFSKRZZVRO-UHFFFAOYSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- OVLZPOPQCGOVIT-UHFFFAOYSA-N C(C)[SiH3].[O] Chemical compound C(C)[SiH3].[O] OVLZPOPQCGOVIT-UHFFFAOYSA-N 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
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- 150000001261 hydroxy acids Chemical group 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
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- 239000003921 oil Substances 0.000 description 1
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- 238000007254 oxidation reaction Methods 0.000 description 1
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- 229920001282 polysaccharide Polymers 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention provides a method for quickly immobilizing proteins or amino-contained molecules on an aminated surface. Amino-contained molecules are firstly assembled on a solid-phase surface such asglass, silicon chip, gold and the like to form an aminated surface, a mixture of N-hydroxy-succinamide and 1-ethide-3-(3-dimethyllaminopropyl) carbodiimide is used for functionalizing the carboxyl ofa poly-carboxyl compound, the functionalized poly-carboxyl compound reacts with the aminated surface, the immobilization of the poly-carboxyl compound on the solid-phase surface is realized by the combination of the functionalized carboxyl and the amino, and the remaining functionalized carboxyl can react with protein molecules to be immobilized and other amino-containing molecules. The method canquickly introduce the functionalized carboxyl to the aminated surface, realize the immobilization of the proteins, simplify surface-modifying steps and shorten the time for immobilizing the proteins.The method has the advantages of simple operation and environment friendliness and is suitable for the covalent immobilization of the amino-contained molecules on the aminated surface.
Description
Technical field
The present invention relates to a kind ofly contain the method for amino molecule, belong to surface chemistry and proteopexy technical field at amino modified solid phase surface ankyrin or other.
Background technology
At present, albumen has multiple at the fixing means of solid chip surface, comprise physisorphtion, chemical covalent method, polymkeric substance entrapping method etc., wherein chemical covalent method ankyrin molecule, have easy and simple to handle, become the firm characteristics of key.Utilize the amino on protein molecular surface, by chemical reaction, with the protein molecular Covalent Immobilization at the modification chip surface: comprising aldehyde grouping modified surface, carboxylated modified surface and epoxy silane modified surface etc.
Aldehyde radicalization and carboxylated two kinds of modified surfaces all are to be terminal silicane molecule (as aminopropyl trimethoxy hexasilane (APTMS), aminopropyl three oxygen ethylsilane (APTES), p-aminophenyl Trimethoxy silane (APhS) etc.) with amino, on the decorative layer basis for self-assembly, carry out further chemical treatment again, finally obtain having the surface of activity functional groups.
Wherein, adopt glutaraldehyde to handle amino modified surface, can obtain the aldehyde radical modified surface, by aldehyde radical and proteic amino western Buddhist alkali (Schiff ' s base), the final ankyrin molecule of forming, though being utilized, this method produces commercial product, but because this process reaction time is longer, generally need to react at least under the room temperature 1 hour, and the western Buddhist alkali that forms is stable inadequately, hydrolysis takes place easily, needs SODIUM CYANO BOROHYDRIDE that it is reduced; Also there is the terephthalaldehyde of employing to replace glutaraldehyde and amino reaction, can strengthens the stability of the western Buddhist alkali that forms.But these methods waste time and energy; And wherein toxicity such as the glutaraldehyde that is adopted, terephthalaldehyde are bigger; Required SODIUM CYANO BOROHYDRIDE reductive agent it be unclear that proteic influence, itself also is difficult for preserving and life-time service.In addition, the silicon chip of aldehyde radical modification is because easily oxidation by air needs special preservation condition, and unsuitable prolonged preservation, so still there are many defectives in the aldehyde radical modified surface.
Carboxy-modified surface then is by succinyl oxide and the reaction of amino modified silicon chip surface, form hydroxy-acid group at silicon chip surface, when the needs ankyrin, can handle carboxy-modified surface by N monohydroxy succinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), carboxyl after the functionalization, can form stable amido linkage with proteic amino rapidly, this reaction promptly is the condensation reaction between the active site of protein amino acid.Adopt this reaction, can shorten the set time of albumen on the surface.Though carboxy-modified surface is relatively stable than the aldehyde radical modified surface, can comparatively make things convenient for preservation with the long period, but this method is handled required time long (this step of only carboxylated modification needs more than 3 hours) in earlier stage, and the efficient of ankyrin is inferior to the aldehyde radical surface.In addition, this method is owing to adopt succinyl oxide to handle amino surface, and reaction process is slow, and the density of uncontrollable chip surface activated carboxyl, and the fixed efficiency of some protein molecular is relevant with the size of carboxyl density.
Summary of the invention
The present invention seeks to overcome the deficiency that prior art exists, provide a kind of and contain the method for amino molecule at the surperficial ankyrin molecule that contains amino or other, be intended to effectively avoid adopting deleterious volatile matter such as aldehydes, reduce environmental pollution, simplify the treatment step on carboxy-modified surface, shorten the proteopexy time.
Purpose of the present invention is achieved through the following technical solutions:
Amino surface quickly immobilizing proteins or contain the method for amino molecule, characteristics are: at first assembling contains amino molecule on solid phase surface, forms amidized surface; Make the carboxyl-functional of multi-carboxy compound with N-hydroxy-succinamide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide mixed solution, with multi-carboxy compound after the functionalization and amidized surface reaction, the multi-carboxy compound of functionalization is attached on the amidized surface, remaining functionalization carboxyl with treat the fixed protein molecular or contain amino compound to react.
Further, above-mentioned amino surface quickly immobilizing proteins or contain the method for amino molecule, described solid phase surface is the surface of glass or silicon chip or gold etc.
Further, above-mentioned amino surface quickly immobilizing proteins or contain the method for amino molecule, described mixed solution with N-hydroxy-succinamide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide makes the carboxyl-functional of multi-carboxy compound, and its multi-carboxy compound is oxalic acid, propanedioic acid, succsinic acid, pentanedioic acid, citric acid, tartrate, ethylenediamine tetraacetic acid (EDTA) or the like.
Further, above-mentioned amino surface quickly immobilizing proteins or contain the method for amino molecule, the final concentration of described N-hydroxy-succinamide is 50mM.
Again further, above-mentioned amino surface quickly immobilizing proteins or contain the method for amino molecule, the final concentration of described 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide is 200mM.
Again further, above-mentioned amino surface quickly immobilizing proteins or contain the method for amino molecule, multi-carboxy compound after the described functionalization and amidized surface reaction time are 5～10 minutes.
Again further, above-mentioned amino surface quickly immobilizing proteins or contain the method for amino molecule, described to contain amino compound be thanomin, quadrol, trimeric cyanamide, benzaminic acid, Tutofusin tris, glucosamine, polyacrylamide, hyaluronic acid, chitosan or the like.
Substantive distinguishing features and obvious improvement that technical solution of the present invention is outstanding are mainly reflected in:
The inventive method can be implemented in carboxyl and proteic the fixing that amino surface is introduced functionalization fast, simplifies the surface modification step, shortens the protecpectic time; Avoid in the surface modification process, using deleterious aldehyde material, help environmental protection; This method also is applicable on amino modified surface, fixes other and contains amino molecule; Also can be when the functionalization multi-carboxy compound, by adding a certain amount of hydroxyl carboxylic acid compound,,, regulate surperficial carboxyl density thereby reach with its carboxyl functionalization together as oxyacetic acid, contain the purpose of amino molecule fixed effect with control albumen or other.This method operation steps simple and fast, and it is environmentally friendly, effectively avoid adopting deleterious volatile matter such as aldehydes, obviously reduce environmental pollution, simplify the treatment step on carboxy-modified surface, shorten the proteopexy time greatly, be fit to the various Covalent Immobilization that contain amino molecule at amino surface, be rated as new technology with novelty, creativeness, practicality.
Description of drawings
Below in conjunction with accompanying drawing technical solution of the present invention is described further:
Fig. 1: reaction process synoptic diagram of the present invention;
Fig. 2: the amino modified surface of the present invention is immunoglobulin (Ig) (IgG) albumen and in conjunction with the ellipse inclined to one side detection gray-scale map behind its antibody fixedly;
Fig. 3: the atomic force microscope figure of the amino modified surperficial set casing glycan of the present invention.
Embodiment
The present invention proposes to contain the method for amino molecule at the surperficial ankyrin molecule that contains amino or other, only needs carry out conventional surface amination modification to glass or silicon chip etc., and amination modified surface is relatively more stable, and it is comparatively convenient to preserve.Need to wait ankyrin or other to contain the amino period of the day from 11 p.m. to 1 a.m that divides, certain is not contained amino multi-carboxy compound carry out functionalization, multi-carboxy compound after the functionalization, a part of carboxyl wherein can be attached to amino surface, another part then can contain amino molecule with the amino of protein molecular or other and form chemical bond, contains amino molecule fixing at amino surface thereby finish albumen or other.
Multi-carboxy compound comprises the compound of two carboxyls at least, preferred oxalic acid, propanedioic acid, succsinic acid, pentanedioic acid, citric acid, tartrate, ethylenediamine tetraacetic acid (EDTA).
The functionalized reagent of carboxyl is a N-hydroxy-succinamide (NHS) and the mixing solutions of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), also can be that other can make behind the carboxyl-functional material with the amino reaction.
As shown in Figure 1, concrete processing step is: assemble the molecule that contains amino earlier on as solid phase surfaces such as glass, silicon chip or gold, form amidized surface; Make the carboxyl-functional of multi-carboxy compound with the NHS/EDC mixed solution, with the multi-carboxy compound after the functionalization and amination surface reaction number minute, so that the multi-carboxy compound of functionalization is attached on the amino surface, remaining then functionalization carboxyl can rapidly and treat that fixed protein molecular or other contain amino molecule and react.
Embodiment 1: amination silicon chip surface ankyrin
Ordinary method is carried out the amination modified of silicon chip surface, specific as follows: with the silicon chip that cleans up with the vitriol oil, hydrogen peroxide (volume ratio 3: 1) oscillation treatment 30 minutes, after water cleans, change over to and contained in 10% aminopropyltriethoxywerene werene (APTES) ethanolic soln oscillation treatment 2 hours, after rinsing well, promptly get amino modified surface.The succsinic acid aqueous solution of preparation 10mM, then NHS and EDC are dissolved in this solution, make the final concentration of NHE and EDC be respectively 50mM and 200mM, with mixing solutions elder generation and amino modified surface reaction 5 minutes, after washing nitrogen dries up, got final product in 10 minutes with treating the reaction of fixed protein solution again, in order to detect the fixed protein molecular, can be again and its antibody response, to strengthen detection signal.This embodiment, the fixed effect that detects amino surface with ellipse polarisation imaging system is good, as the fixing IgG albumen and in the amino modified surface of Fig. 2 in conjunction with the ellipse inclined to one side detection gray-scale map behind the anti-IgG, at the bottom of being silicon wafer-based from left to right, fixedly IgG albumen, again in conjunction with the grey scale change behind the anti-IgG, gray-scale value is corresponding with the molecule area density on surface, as seen, satisfy the detection requirement.
Embodiment 2: amination silicon chip surface set casing glycan
Ordinary method is carried out the amination modified of silicon chip surface, the citric acid of preparation 10mM, then NHS and EDC are dissolved in this solution, both final concentrations are 50mM/200mM, with this mixing solutions elder generation and amino modified surface reaction 5 minutes, after washing, nitrogen dry up, got final product in 10 minutes with chitosan (containing amino polysaccharide molecule) solution reaction again.
This embodiment is by the atomic force microscope analysis, and the surface is fixed with the higher particulate state chitosan molecule of density really, the atomic force microscope figure of amino modified as shown in Figure 3 surperficial set casing glycan, and the surface particles thing is chitosan molecule.
In sum, the present invention a kind ofly contains the method for amino molecule at amino modified solid surface ankyrin or other, promptly amino modified solid surface utilize functionalization multi-carboxy compound, albumen or other contained amino molecule be fixed on the surface.Effective ankyrin or other contain amino molecule, and can be by the mono carboxylic compound of functionalization, and adjusting albumen or other contain the constant density of amino molecule.This method operation steps simple and fast, and environmentally friendly, be fit to the various Covalent Immobilization that contain amino molecule at amino surface.Compared with prior art, effectively avoid adopting deleterious volatile matter such as aldehydes, obviously reduce environmental pollution, simplify the treatment step on carboxy-modified surface, shorten the proteopexy time greatly, its application prospect is very wide.
Below only be concrete exemplary applications of the present invention, protection scope of the present invention is not constituted any limitation.All employing equivalents or equivalence are replaced and the technical scheme of formation, all drop within the rights protection scope of the present invention.
Claims (7)
1. amino surface quickly immobilizing proteins or contain the method for amino molecule, it is characterized in that: at first assembling contains amino molecule on solid phase surface, forms amidized surface; Make the carboxyl-functional of multi-carboxy compound with N monohydroxy succinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide mixed solution, with multi-carboxy compound after the functionalization and amidized surface reaction, the multi-carboxy compound of functionalization is attached on the amidized surface, remaining functionalization carboxyl with treat the fixed protein molecular or contain amino compound to react.
2. amino surface quickly immobilizing proteins according to claim 1 or contain the method for amino molecule, it is characterized in that: described solid phase surface is glass surface or silicon chip surface or gold surface.
3. amino surface quickly immobilizing proteins according to claim 1 or contain the method for amino molecule, it is characterized in that: described mixed solution with N-hydroxy-succinamide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide makes the carboxyl-functional of multi-carboxy compound, and its multi-carboxy compound is a kind of in oxalic acid, propanedioic acid, succsinic acid, pentanedioic acid, citric acid, tartrate, the ethylenediamine tetraacetic acid (EDTA).
4. amino surface quickly immobilizing proteins according to claim 1 or contain the method for amino molecule, it is characterized in that: the final concentration of described N-hydroxy-succinamide is 50mM.
5. amino surface quickly immobilizing proteins according to claim 1 or contain the method for amino molecule, it is characterized in that: the final concentration of described 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide is 200mM.
6. amino surface quickly immobilizing proteins according to claim 1 or contain the method for amino molecule, it is characterized in that: multi-carboxy compound after the described functionalization and amidized surface reaction time are 5～10 minutes.
7. amino surface quickly immobilizing proteins according to claim 1 or contain the method for amino molecule is characterized in that: described to contain amino compound be a kind of in thanomin, quadrol, trimeric cyanamide, benzaminic acid, Tutofusin tris, glucosamine, polyacrylamide, hyaluronic acid, the chitosan.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106729941A (en) * | 2017-03-07 | 2017-05-31 | 南京林业大学 | A kind of preparation method of chitosan-based sponge dressing |
| CN110551303A (en) * | 2019-09-17 | 2019-12-10 | 江南大学 | Method for preparing flexible conductive silk fibroin membrane by enzyme method |
| WO2021224833A1 (en) * | 2020-05-06 | 2021-11-11 | Universidad De Los Andes | Bio-nanocompound as an agent for nucleating aqueous-based compounds and production method thereof |
-
2009
- 2009-05-12 CN CN200910027564A patent/CN101671395A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106729941A (en) * | 2017-03-07 | 2017-05-31 | 南京林业大学 | A kind of preparation method of chitosan-based sponge dressing |
| CN110551303A (en) * | 2019-09-17 | 2019-12-10 | 江南大学 | Method for preparing flexible conductive silk fibroin membrane by enzyme method |
| CN110551303B (en) * | 2019-09-17 | 2021-08-17 | 江南大学 | Method for preparing flexible conductive silk fibroin membrane by enzyme method |
| WO2021224833A1 (en) * | 2020-05-06 | 2021-11-11 | Universidad De Los Andes | Bio-nanocompound as an agent for nucleating aqueous-based compounds and production method thereof |
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