CN1046549C - Method for biological sensor produced by latex embedding enzyme - Google Patents
Method for biological sensor produced by latex embedding enzyme Download PDFInfo
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- CN1046549C CN1046549C CN96123528A CN96123528A CN1046549C CN 1046549 C CN1046549 C CN 1046549C CN 96123528 A CN96123528 A CN 96123528A CN 96123528 A CN96123528 A CN 96123528A CN 1046549 C CN1046549 C CN 1046549C
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- enzyme
- biological sensor
- latex
- embedding
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- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention belongs to a method for preparing a biological sensor by embedding an enzyme by latex, which aims to provide the method for preparing a biological sensor by embedding an enzyme by latex. In the method for preparing a biological sensor by embedding an enzyme by latex, after the enzyme is dissolved in the water solution of polyvinyl alcohol grafting 4-vinylpyridine or the mixed liquid of dimethyl sulfoxide and water, the obtained mixed liquid is directly dripped and coated on an electrode surface, a placing process lasts for a period of time at low temperature, and the biological sensor with an enzyme membrane firmly stuck to the electrode surface is obtained. The biological sensor prepared by the method for preparing a biological sensor by embedding an enzyme by latex has the advantages of short balancing time, fast response, good reproducibility and long service life.
Description
The invention belongs to the method for biological sensor produced by latex embedding enzyme.
Immobilized enzyme is the committed step of preparation biosensor.Up to now, using the most general immobilization technology is to adopt gel/polymkeric substance embedding.It can and be fixed in the high-polymer three-dimensional spacial framework enzyme molecule or cell embedding.B.A.Gregg, A.Heller, J.Phys.Chem., 1991,95,5976 disclose a kind of at electrode surface with the long-chain bis-epoxy crosslinked poly 4 vinyl pyridine and the method for immobilized enzyme simultaneously, gained contain the enzyme hydrogel can secure adhesion in electrode surface and response is faster arranged, but pure hydrophobic 4-vinylpridine carrier framework is unfavorable for the biocompatibility and the biologically stable of enzyme, and bis-epoxy chemically crosslinked process can cause a small amount of enzyme deactivation.Tongyin Yu, Haiying Liu, Jiaqi Deng and Yongcheng Liu, at J.of Appl.Polym.Sci., 1995,58, reported a kind of method for preparing biosensor with regenerated silk embedding enzyme in 973, its biggest advantage is to have avoided chemically crosslinked or radical polymerization etc. to cause the operation of enzyme deactivation easily in the immobilized enzyme process, but the character by silk itself is determined, the glued membrane that this method makes is dry or can become fragile when being exposed in the air for a long time, and then influences the performance of biosensor.
The method that the purpose of this invention is to provide a kind of biological sensor produced by latex embedding enzyme, after soon enzyme will be dissolved in the aqueous solution or dimethyl sulfoxide (DMSO)/water mixed liquid of polyvinyl alcohol graft copolymerized 4-vinyl pyridine, direct the dripping of the mixed liquid of gained is applied to electrode surface, low temperature is placed for some time, make polyvinyl alcohol graft copolymerized 4-vinyl pyridine self physical crosslinking become glue and the enzyme of embedding simultaneously, obtain the biosensor of enzyme membrane secure adhesion in electrode surface, the biosensor starting time of preparing like this is short, response is fast, favorable reproducibility, long service life.
Polyvinyl alcohol is a kind of polyhydroxylated polymer of synthetic, have excellent biological compatibility and biologically stable, can be in water or dimethyl sulfoxide (DMSO)/water mixed liquid become glue and the enzyme of embedding simultaneously by self physical crosslinking, and gained contains enzyme glued membrane stability and wearing quality is better, thereby is widely used as the fixed enzyme vector in biosensor and the bio-reactor.The present invention selects for use the 4-vinylpridine grafts of polyvinyl alcohol as fixed enzyme vector, when having kept all advantages of polyvinyl alcohol, has greatly increased the adhesive power of enzyme membrane at electrode surface, and simultaneously, the effect of immobilized enzyme is also better.
It is 2～5% the aqueous solution or dimethyl sulfoxide (DMSO)/water mixed liquid that the present invention is made into concentration with the 4-vinylpridine grafts of polyvinyl alcohol, again with 0.5～3.0 milligram enzyme, these enzymes are glucose oxidase or horseradish peroxidase or polyphenoloxidase, add in this solution of 150～300 microlitres, also can be with 1 * 10
-2MolL
-1The mediator yellow prussiate of potash add simultaneously wherein, after mixing, this mixing drop of 5～15 microlitres is coated onto the basal electrode surface, placed naturally 16～24 hours, and promptly got required biosensor for-20 ℃～+ 4 ℃.This method also can be used for alcohol oxidase, rCO, alcoholdehydrogenase, laccase, cholinesterase, E.C. 1.1.99.1, bilirubin oxidase, lipase fixing.
The preparation method of biosensor of the present invention has selected a kind of novel fixed enzyme vector for use--the 4-vinylpridine grafts of polyvinyl alcohol, it has excellent biological compatibility and biologically stable, and can with the electrode surface mortise; Become glue and the enzyme of embedding simultaneously by carrier self physical crosslinking, the enzyme that can avoid chemical cross-linking agent or free radical etc. to cause is lived and is lost; Can make hydrogel or dimethyl sulfoxide (DMSO)/water mixed liquid organogel according to the needs of detection architecture, be respectively applied for the detection in water and the organic phase; The preparation method is simple, favorable reproducibility; The biosensor that makes is applied widely, and starting time is short, and response is fast, good stability, long service life.
Embodiment provided by the invention is as follows:
Embodiment 1: glucose oxidase electrode.3 milligrams of glucose oxidases are dissolved in the polyvinyl alcohol graft copolymerized 4-vinyl pyridine aqueous solution of 150 microlitres 2%, after stirring, pipette this drips of solution of 6 microlitres with microsyringe and be applied to platinum electrode surface ,+4 ℃ of refrigerators were placed 24 hours, promptly made glucose oxidase electrode.This electrode can be used for aqueous phase and detects glucose, electrode starting time 1～2 minute; 5～8 seconds time of response; Linearity range 5 * 10
-6～5 * 10
-3Mo1L
-1Detectability 2 * 10
-6Mo1L
-1Stability, response value is about 85% of an initial value after 2 months.
Embodiment 2: the horseradish peroxidase electrode.0.6 milligram of horseradish peroxidase is dissolved in the polyvinyl alcohol graft copolymerized 4-vinyl pyridine aqueous solution of 200 microlitres 5%, after stirring, pipette this drips of solution of 10 microlitres with microsyringe and be applied to graphite electrode surface, 0 ℃ of refrigerator was placed 20 hours, promptly made the horseradish peroxidase electrode.This electrode can be used for the detection of materials such as aqueous phase hydrogen peroxide, phenol, amine, in the electrode starting time 3 minutes; 10～15 seconds time of response; Stability is more than 2 months.
Embodiment 3: Polyphenol Oxidase Electrode.0.5 milligram of polyphenoloxidase is dissolved in the polyvinyl alcohol graft copolymerized 4-vinyl pyridine aqueous solution of 300 microlitres 5%, after stirring, pipette this drips of solution of 8 microlitres with microsyringe and be applied to the glass-carbon electrode surface ,-10 ℃ of refrigerators were placed 18 hours, promptly made Polyphenol Oxidase Electrode.This electrode can be measured materials such as phenol, catechol at aqueous phase.In the electrode starting time 3 minutes; 20～25 seconds time of response; Stability is more than 2 months.
Embodiment 4: organic phase horseradish peroxidase electrode.0.8 milligram of horseradish peroxidase is dissolved in the dimethyl sulfoxide (DMSO)/water mixed liquid of polyvinyl alcohol graft copolymerized 4-vinyl pyridine of 200 microlitres 4%, wherein dimethyl sulfoxide (DMSO) accounts for 40% of cumulative volume, and adding yellow prussiate of potash again and making its ultimate density is 1 * 10
-2MolL
-1, after stirring, pipetting this drips of solution of 12 microlitres with microsyringe and be applied to the glass-carbon electrode surface ,-20 ℃ of refrigerators were placed 16 hours, promptly made organic phase horseradish peroxidase electrode.This electrode can be used for measuring in the organic solvents such as chloroform the detection of materials such as hydrogen peroxide, organo-peroxide and phenol, amine, in the electrode starting time 10 minutes; 60～90 seconds time of response; Stability is more than 2 months.
Claims (2)
1. a gel embedding prepares the method for biosensor, it is characterized in that it is 2～5% the aqueous solution or dimethyl sulfoxide (DMSO)/water mixed liquid that 4-vinylpridine grafts with polyvinyl alcohol is made into concentration, again with 0.5～3.0 milligram enzyme, these enzymes are glucose oxidase or horseradish peroxidase or polyphenoloxidase, add in this solution of 150～300 microlitres, perhaps further with 1 * 10
-2MolL
-1Mediator add simultaneously wherein, after mixing, this mixing drop of 5～15 microlitres is coated onto the basal electrode surface, placed naturally 16～24 hours, and promptly got required biosensor for-20 ℃～+ 4 ℃.
2. the method for preparing biosensor as claimed in claim 1 is characterized in that described mediator is a yellow prussiate of potash.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN96123528A CN1046549C (en) | 1996-12-25 | 1996-12-25 | Method for biological sensor produced by latex embedding enzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN96123528A CN1046549C (en) | 1996-12-25 | 1996-12-25 | Method for biological sensor produced by latex embedding enzyme |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1186115A CN1186115A (en) | 1998-07-01 |
| CN1046549C true CN1046549C (en) | 1999-11-17 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN96123528A Expired - Fee Related CN1046549C (en) | 1996-12-25 | 1996-12-25 | Method for biological sensor produced by latex embedding enzyme |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1046549C (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1061372C (en) * | 1997-10-10 | 2001-01-31 | 中国科学院感光化学研究所 | Oxidase function compound sensitive film containing hydrophobic nanometre prill and its making method and use |
| JP3694424B2 (en) * | 1998-09-29 | 2005-09-14 | 松下電器産業株式会社 | Glucose sensor |
| CN101693873B (en) * | 2009-10-15 | 2011-12-28 | 天津大学 | Sandwich structuralized enzymatic membrane reactor, production method and application thereof |
| CN106635931B (en) * | 2017-02-17 | 2019-09-27 | 青岛中科煜成安全技术有限公司 | Engineering bacteria for BOD biosensor |
| CN107760665B (en) * | 2017-09-29 | 2020-05-08 | 浙江大学 | Hydrogel-wrapped single cell-based method and product and application in preparation of universal red blood cells |
| CN110220877A (en) * | 2019-06-12 | 2019-09-10 | 万细凤 | Based on SiO2The novel glucose composite sensitive film and preparation method of nanoparticle |
| CN110257028B (en) * | 2019-06-17 | 2022-04-05 | 同济大学 | Enzymatic polymerization macromolecule composite slurry and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS55124060A (en) * | 1979-03-16 | 1980-09-24 | Matsushita Electric Ind Co Ltd | Enzyme electrode |
| CN85107223A (en) * | 1984-12-17 | 1986-07-09 | 株式会社岛津制作所 | Enzyme electrode |
| US4894253A (en) * | 1986-08-12 | 1990-01-16 | University Of Cincinnati | Method for production of coated electrode |
| CN1076488A (en) * | 1992-03-17 | 1993-09-22 | 陈国诚 | The preparation method of polyvinyl alcohol microbe or enzyme immobilization carrier and application thereof |
| JPH06109692A (en) * | 1992-09-25 | 1994-04-22 | A & D Co Ltd | Oxygen electrode |
| CN1101943A (en) * | 1994-08-26 | 1995-04-26 | 北京市食品酿造研究所 | Process for preparing immobilized living cells and enzyme |
-
1996
- 1996-12-25 CN CN96123528A patent/CN1046549C/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS55124060A (en) * | 1979-03-16 | 1980-09-24 | Matsushita Electric Ind Co Ltd | Enzyme electrode |
| CN85107223A (en) * | 1984-12-17 | 1986-07-09 | 株式会社岛津制作所 | Enzyme electrode |
| US4894253A (en) * | 1986-08-12 | 1990-01-16 | University Of Cincinnati | Method for production of coated electrode |
| CN1076488A (en) * | 1992-03-17 | 1993-09-22 | 陈国诚 | The preparation method of polyvinyl alcohol microbe or enzyme immobilization carrier and application thereof |
| JPH06109692A (en) * | 1992-09-25 | 1994-04-22 | A & D Co Ltd | Oxygen electrode |
| CN1101943A (en) * | 1994-08-26 | 1995-04-26 | 北京市食品酿造研究所 | Process for preparing immobilized living cells and enzyme |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1186115A (en) | 1998-07-01 |
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