CN1346983A - Process for preparing biologic sensor by embedding enzyme with composite titanium oxide sol-gel membrane - Google Patents
Process for preparing biologic sensor by embedding enzyme with composite titanium oxide sol-gel membrane Download PDFInfo
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- CN1346983A CN1346983A CN 01128285 CN01128285A CN1346983A CN 1346983 A CN1346983 A CN 1346983A CN 01128285 CN01128285 CN 01128285 CN 01128285 A CN01128285 A CN 01128285A CN 1346983 A CN1346983 A CN 1346983A
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Abstract
A process for preparing biological sensor by embedding enzyme in composite titanium oxide sol-gel film includes hydrolyzing the titanium isopropoxide to obtain titanium oxide sol, mixing it with the enzyme-containing polyvinyl alcohol grafted 4-vinyl pyridine solution, directly applying the mixture on the surface of electrode, and standing at 4 deg.C for 24-48 hr to obtain a biological sensor. Its advantages are high adhesion to electrode, high response speed and sensitivity and long service life.
Description
Technical field: the invention belongs to the method that embedding enzyme with composite titanium oxide sol-gel membrane prepares biology sensor.
Background technology: the sol-gel investment is a kind of effective immobilised enzymes method that developed recently gets up, utilize this technology can be at low temperatures to the alkoxide hydrolysis, concentrate and can form network-like film, has biocompatibility, be very suitable for the fixing of biomolecule, and the enzyme glued membrane that contains that makes has physics rigidity, advantages such as chemical inertness and insignificant swelling.The biology sensor of the sol gel film immobilized enzyme of having reported at present mainly concentrates on silane systems, and Deng family is auspicious in addition waits the people to report for work with alundum (Al sol gel film immobilized enzyme, but the film of above-mentioned two kinds of immobilized enzymes all is nonconducting.In order to accelerate enzyme molecule and interelectrode electron transfer, Glezer and Lev is at J.Am.Chem.Soc.1993,115, delivered a kind of method for preparing biology sensor with vanadium pentoxide gel entrapment glucose oxidase among the 2533-2534, because tetravalent vanadium ion has good electron conduction, cause this film to have semi-conductive character, be applicable to the support material of reactive protein, it is reported that the biology sensor that makes with this method is active in 10 days when storing for 4 ℃ does not reduce.
Summary of the invention: the purpose of this invention is to provide the new method that a kind of embedding enzyme with composite titanium oxide sol-gel membrane prepares biology sensor, be about to titanium isopropoxide hydrolysis TiO 2 sol that makes and the polyvinyl alcohol graft copolymerized 4-vinyl pyridine aqueous solution that contains enzyme, directly drip behind the mixing and be applied to electrode surface, placed 24-48 hour down for 4 ℃, obtain containing the even glued membrane of enzyme and adhere to electrode surface securely, the biology sensor of preparing like this is highly sensitive, response is fast, can maintain more than one month serviceable life, is suitable for detecting various zymolytes.
The present invention is dissolved in 20-40 μ l titanium isopropoxide in the isopropyl alcohol of 10-15 μ l, adds 7-14 μ l0.1mol L
-1Hydrochloric acid and 240-480 μ l aqueous solution, mixing, sonic oscillation got Sol A after 1 hour; Be that 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine is made into the aqueous solution that concentration is 8-10% (w/v) with grafting degree in addition, again with the enzyme of 1-3 milligram, promptly a kind of in glucose oxidase, horseradish peroxidase, the polyphenol oxidase adds in the above-mentioned aqueous solution of 100 μ l, mixing gets solution B; Then with Sol A and solution B in 1: the ratio mixing of 1-2.5, pipette this mixed liquor 5-10 μ l with micro syringe and drip and be coated onto the basal electrode surface, placed 24-48 hour down, promptly make required biology sensor for 4 ℃.
Compare with the method for traditional preparing bio-sensor with embedding enzyme of collosol-gel, the method for preparing biology sensor that the present invention proposes has many advantages.On the one hand titania is a kind of semiconductor material, helps promoting the electron transfer between enzyme molecule and the electrode; TiO 2 particles has excellent biological compatibility simultaneously, helps at utmost keeping biomolecule activity; Amido, carboxyl and the hydroxyl of titanium and enzyme molecular surface have coordination in addition, help the fixing of enzyme molecule; Polyvinyl alcohol graft copolymerized 4-vinyl pyridine can be by interaction stabilized enzyme such as hydrogen bonds on the other hand, and the 4-vinylpridine side chain all has stronger suction-operated at multiple electrode surface simultaneously, can strengthen to contain the adhesion of enzyme glued membrane at electrode surface.When titanium oxide colloid is mixed mutually with polyvinyl alcohol graft copolymerized 4-vinyl pyridine, can form interpenetrating networks, make firm being fixed in the composite membrane of enzyme molecule.The character of this composite membrane is better than using separately each Substance Properties.
Embodiment is as follows:
Embodiment 1:
Glucose oxidase sensor.30 μ l titanium isopropoxides are dissolved in the isopropyl alcohol of 13 μ l, add 360 μ l water and 10 μ l 0.1mol L
-1Hydrochloric acid mixed solution, mixing, sonic oscillation got Sol A after 1 hour; In addition be with grafting degree 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine to be made into concentration be 8% aqueous solution, in 70 these solution of μ l, add 2 milligrams glucose oxidase, mixing, solution B; Then with Sol A and solution B ratio mixing in 1: 2.5, pipette 8 μ l with micro syringe and should mix drop and be coated onto the glass-carbon electrode surface, placed 24 hours down for 4 ℃, promptly make glucose biological sensor, can be used for aqueous phase and detect glucose.This biology sensor equilibration time is in 6 minutes; Response time is 15 seconds; The range of linearity is 5 * 10
-6-9 * 10
-3Mol L
-1Stability keeps more than 1 month.
Embodiment 2:
The polyphenol oxidase enzyme biologic sensor.20 μ l titanium isopropoxides are dissolved in the 10 μ l isopropyl alcohols, add 7 μ 10.1mol L
-1Hydrochloric acid and 240 μ l aqueous solution, mixing, sonic oscillation got Sol A after 1 hour; In addition be with grafting degree 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine to be made into concentration be 10% aqueous solution, in 100 these solution of μ l, add 1.5 milligrams polyphenol oxidase mixing, solution B; Then with Sol A and solution B ratio mixing in 1: 1, pipette 10 μ l with micro syringe and should mix drop and be coated onto the glass-carbon electrode surface, placed 30 hours down, promptly make the polyphenol oxidase enzyme biologic sensor for 4 ℃, can be used for aqueous phase and measure phenol, materials such as catechol.This biology sensor equilibration time is in 5 minutes; Response time is 15 seconds; Stability keeps more than 2 months.
Embodiment 3:
The horseradish peroxidase sensor.40 μ l titanium isopropoxides are dissolved in the 15 μ l isopropyl alcohols, add 14 μ l 0.1mol L
-1Hydrochloric acid and 480 μ l aqueous solution, mixing, sonic oscillation got Sol A after 1 hour; In addition be with grafting degree 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine to be made into concentration be 9% aqueous solution, in 100 these solution of μ l, add 1 milligram horseradish peroxidase, mixing, solution B; Then with Sol A and solution B ratio mixing in 1: 2, pipette 5 μ l with micro syringe and should mix drop and be applied to graphite electrode surface, placed 48 hours down for 4 ℃, promptly make the horseradish peroxidase sensor, can be used for aqueous phase and detect hydrogen peroxide, phenol, materials such as amine.This biology sensor equilibration time is in 5 minutes; Response time is 15 seconds; Stability keeps more than 2 months.
Claims (4)
1. an embedding enzyme with composite titanium oxide sol-gel membrane prepares the method for biology sensor, it is characterized in that 20-40 μ l titanium isopropoxide is dissolved in the 10-15 μ l isopropyl alcohol, adds 7-14 μ l0.1mol L
-1Hydrochloric acid and 240-480 μ l aqueous solution, mixing, sonic oscillation got Sol A after 1 hour; Other gets grafting degree is that 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine is made into the aqueous solution that concentration is 8-10%, a kind of with in the glucose oxidase of 1-3 milligram, polyphenol oxidase, the horseradish peroxidase again, add in the above-mentioned aqueous solution of 100 μ l, mixing gets solution B; Then with Sol A and solution B in 1: the ratio mixing of 1-2.5, pipette 5-10 μ l with micro syringe and should mix drop and be coated onto the basal electrode surface, placed 24-48 hour down, promptly make required biology sensor for 4 ℃.
2. embedding enzyme with composite titanium oxide sol-gel membrane as claimed in claim 1 prepares the method for biology sensor, it is characterized in that getting grafting degree and be 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine and be made into the aqueous solution that concentration is 8-10%, glucose oxidase is added in the above-mentioned aqueous solution again, mixing gets solution B.
3. embedding enzyme with composite titanium oxide sol-gel membrane as claimed in claim 1 prepares the method for biology sensor, it is characterized in that getting grafting degree and be 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine and be made into the aqueous solution that concentration is 8-10%, polyphenol oxidase is added in the above-mentioned aqueous solution again, mixing gets solution B.
4. embedding enzyme with composite titanium oxide sol-gel membrane as claimed in claim 1 prepares the method for biology sensor, it is characterized in that getting grafting degree and be 12.5% polyvinyl alcohol graft copolymerized 4-vinyl pyridine and be made into the aqueous solution that concentration is 8-10%, horseradish peroxidase is added in the above-mentioned aqueous solution again, mixing gets solution B.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB011282851A CN1156692C (en) | 2001-10-12 | 2001-10-12 | Process for preparing biologic sensor by embedding enzyme with composite titanium oxide sol-gel membrane |
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| CNB011282851A CN1156692C (en) | 2001-10-12 | 2001-10-12 | Process for preparing biologic sensor by embedding enzyme with composite titanium oxide sol-gel membrane |
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| CN1346983A true CN1346983A (en) | 2002-05-01 |
| CN1156692C CN1156692C (en) | 2004-07-07 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100385234C (en) * | 2006-04-18 | 2008-04-30 | 湖南大学 | Sensor for detecting phenolic substance in compost and detection method thereof |
| CN101393161B (en) * | 2008-10-29 | 2011-11-02 | 北京化工大学 | Bio-sensing multilayer film of titanium oxide contained nanometer sheet and method for making same |
| CN101281157B (en) * | 2008-04-30 | 2012-06-27 | 中国科学院上海硅酸盐研究所 | Decorating electrode of transient metal to replace laminar titanium oxide fixing protein |
| CN103628052A (en) * | 2013-09-04 | 2014-03-12 | 杭州电子科技大学 | Preparation method of titanium dioxide thin film coating on surface of titanium alloy microporous bone |
| CN108414595A (en) * | 2017-06-18 | 2018-08-17 | 苏州科技大学 | Utilize TiO2The method of glucose in nanometer tube modified ito determination of electrode aqueous solution |
-
2001
- 2001-10-12 CN CNB011282851A patent/CN1156692C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100385234C (en) * | 2006-04-18 | 2008-04-30 | 湖南大学 | Sensor for detecting phenolic substance in compost and detection method thereof |
| CN101281157B (en) * | 2008-04-30 | 2012-06-27 | 中国科学院上海硅酸盐研究所 | Decorating electrode of transient metal to replace laminar titanium oxide fixing protein |
| CN101393161B (en) * | 2008-10-29 | 2011-11-02 | 北京化工大学 | Bio-sensing multilayer film of titanium oxide contained nanometer sheet and method for making same |
| CN103628052A (en) * | 2013-09-04 | 2014-03-12 | 杭州电子科技大学 | Preparation method of titanium dioxide thin film coating on surface of titanium alloy microporous bone |
| CN103628052B (en) * | 2013-09-04 | 2016-01-20 | 杭州电子科技大学 | A kind of preparation method of titanium alloy micropore bone surface titanium deoxid film coating |
| CN108414595A (en) * | 2017-06-18 | 2018-08-17 | 苏州科技大学 | Utilize TiO2The method of glucose in nanometer tube modified ito determination of electrode aqueous solution |
| CN108414595B (en) * | 2017-06-18 | 2020-06-16 | 苏州科技大学 | By using TiO2Method for determining glucose in aqueous solution by nanotube modified ito electrode |
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| Publication number | Publication date |
|---|---|
| CN1156692C (en) | 2004-07-07 |
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