CN1635370A - Method for preparing biosensor - Google Patents
Method for preparing biosensor Download PDFInfo
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- CN1635370A CN1635370A CN 200310117748 CN200310117748A CN1635370A CN 1635370 A CN1635370 A CN 1635370A CN 200310117748 CN200310117748 CN 200310117748 CN 200310117748 A CN200310117748 A CN 200310117748A CN 1635370 A CN1635370 A CN 1635370A
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- haemoglobin
- sensor
- film
- silicon sol
- gel
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Abstract
This invention relates to the silicon sol to gel film hemoglobin loading sensor process method. This invention adopts the thin film by on spot hydrolytic polymerization of silane in alkalescence conditions and fixes the film on the carbon electrode surface to make biological sensor. The sensor has catalyzing current response to auricome, oxygen, nitrite and trichloroacetic acid.
Description
Technical field
The invention belongs to a kind of preparation method of biology sensor, concretely, the invention belongs to the preparation method of the immobilized haemoglobin third generation of silicon sol-gel film biology sensor.
Background technology
Biopotential sensor has in fields such as industrial and agricultural production, health care, environmental protection widely to be used.Biology sensor is to utilize biological substance as recognition component, and concentration and measurable electric signal of measured matter associated.What research was maximum in the biology sensor is enzyme sensor.Mechanism according to electron transfer between enzyme and electrode roughly can be divided into the three generations with enzyme biologic sensor: in glucose sensor, the enzyme sensor that the Catalysis Principles of natural amboceptor one oxygen of employing enzyme designs and produces is called first generation biology sensor; Second generation biology sensor mixes the mediator of synthetic in the enzyme layer, has reduced the interference of space oxygen; Third generation biology sensor is meant in the presence of the medialess body, the enzyme sensor that utilizes enzyme and interelectrode direct electron transfer to make.In recent years, third generation Study on Biosensor is very active, but great majority all concentrate on native enzyme (as horseradish peroxidase HRP) as recognition component.Yet the shortcoming of the third generation biology sensor maximum of native enzyme base is to use the life-span weak point, costs an arm and a leg.
Be expected to replace the recognition component of HRP as biology sensor owing to haemoglobin has the iron porphyrin prothetic group identical with HRP, the research of this respect has attracted the interest of a large amount of chemists.Yet because haemoglobin molecule is huge, the activated centre is buried among folding peptide chain.So, make haemoglobin become the recognition component of third generation biology sensor, the difficulty that need overcome is exactly to realize its direct electron transfer.Just should effective immobilized haemoglobin in electrode surface, prevent again owing to immobilized its sex change inactivation that makes, to realize that also the direct electron between haemoglobin and the electrode shifts.Silicon sol-gel process condition gentleness, chemical inertness, the bioactivator that is embedded in the sol-gel can keep its structure, activity and function, and micromolecule can contact with bioactivator by the duct of sol-gel.Based on this, silicon sol-gel has been widely used in the immobilized biology enzyme of electrode surface and has developed glucose, H
2O
2Sensor etc.
Summary of the invention
The preparation method who the purpose of this invention is to provide the immobilized haemoglobin third generation of a kind of silicon sol-gel film biology sensor.
Our invention thought is to be applied to the carbon paste electrode surface of having modified haemoglobin with the silicon sol-gel storing solution, make its at room temperature on-the-spot base catalyzed polymerization film forming and make haemoglobin immobilized, thereby make the immobilized haemoglobin third generation of silicon sol-gel film biology sensor.The sensor of making has the catalysis characteristics of stronger peroxidase, can be to H
2O
2, O
2, electrical catalyze reduction optionally such as nitrite, trichloroacetic acid.This sensor reaches balance time weak point, fast, simple, cheap, the long service life of making of response.
A kind of preparation method of biology sensor is characterized in that the phosphate solution that this method will contain the 15-20mg/mL haemoglobin is applied to the carbon paste electrode surface, and drying at room temperature generates the haemoglobin film; Evenly be coated with the silicon sol-gel liquid of forming by silane, alcohol, sodium hydroxide solution on haemoglobin film surface again, generate the immobilized haemoglobin biology sensor of silicon sol-gel film through the room temperature base catalyzed polymerization.
The used silane of the present invention is selected from tetraethoxysilane or tetramethoxy-silicane.
The used alcohol of the present invention is selected from ethanol or methyl alcohol.
Because the immobilized haemoglobin of the present invention uses the silicon sol-gel film, and has adopted on-the-spot base catalysis film forming, thereby avoided strong acid to make catalyzer and the sex change of the haemoglobin that causes.Experimental results show that immobilized haemoglobin in the carbon paste electrode surface has kept its original configuration state, thereby shown the catalytic activity of stronger peroxidase.In addition, the sol gel film of silicon has a large amount of oh groups can form hydrogen bond with protein, can prevent that haemoglobin from coming off from electrode surface; Add that haemoglobin itself is more stable than HRP, thereby the third generation biology sensor that makes the present invention make has the serviceable life that stronger operational stability is arranged and grow than biology enzyme.
Because the basal electrode that the present invention adopts is carbon paste electrode, recognition component is that haemoglobin, immobilized haemoglobin use the silicon sol-gel film.Material gets easily, makes simple, cheap.
The biology sensor of the present invention's preparation has soda acid adaptability preferably, can all demonstrate catalytic activity and stability preferably in the scope of pH4-9.
Embodiment
Embodiment 1:
After 5g dag and 3mL whiteruss mixed, get and insert glass tube [4.5mm (i.d.) in right amount; 5.5mm the height of the about 5mm of an end (o.d.)], the other end inject a copper cash and constitute carbon paste electrode.The surface of electrode ground smooth on pan paper.
NaOH solution and 60 μ LH with 600 μ L ethanol, 50 μ L tetraethoxysilanes (TEOS), 10 μ L 5mmol/L
2O obtains silicon sol-gel liquid after the rapid mixing in a small test tube, it is standby to place 4 ℃ of refrigerators to preserve.
The phosphate buffered solution (pH7.0) of 5 μ L 15mg/mL haemoglobins is dripped to the surface of carbon paste electrode, evenly with the kapillary coating of an end closure; Room temperature airing 1h.Then, 10 μ L silicon sol-gel storing solutions are dripped to the electrode surface of having been modified by Hb also be coated with evenly, make its polymerization at room temperature 1 hour.At last, the electrode (that is: the immobilized haemoglobin third generation of silicon sol-gel film biology sensor) for preparing is dipped in places 4 ℃ of refrigerators to preserve in the phosphate buffered solution of pH7.0.This sensor has good response performance: it is short to reach equilibration time, is not more than 10 minutes; Response is fast, about 0.5-1 minute; Long service life is more than 10 weeks.
Embodiment 2:
Consisting of of silicon sol-gel liquid: NaOH solution and the 60 μ LH of 600 μ L methyl alcohol, 50 μ L tetramethoxy-silicanes (TMOS), 10 μ L 5mmol/L
2O.Other operation steps is with embodiment 1.
The performance of the sensor of embodiment 1 and embodiment 2 preparations:
Sensor is used for the aqueous solution determination of peroxide
The hydrogen peroxide response sensitivity is: 64.8 μ A/mmol/L.
Detect and be limited to: 8.6 * 10
-7Mol/L
The measure linear scope is 1 * 10
-6-2.8 * 10
-4Mol/L.
Sensor is used for the mensuration of aqueous solution nitrite
Detect and be limited to: 3 * 10
-7Mol/L.
The measure linear scope is: 0-8.6 * 10
-4Mol/L.
Sensor is used for the mensuration of aqueous solution trichloroacetic acid
Detect and be limited to: 3 * 10
-6Mol/L.
The measure linear scope is 2.3 * 10
-5-8.6 * 10
-3Mol/L.
Sensor is used for the mensuration of aqueous solution dissolved oxygen DO under 22 ℃ room temperature condition
Detect and be limited to: 5.2 * 10
-6Mol/L
The measure linear scope is: 7.8 * 10
-6-1 * 10
-3Mol/L.
Claims (3)
1, a kind of preparation method of biology sensor is characterized in that the phosphate solution that this method will contain the 15-20mg/mL haemoglobin is applied to the carbon paste electrode surface, and drying at room temperature generates the haemoglobin film; Evenly be coated with the silicon sol-gel liquid of forming by silane, alcohol, sodium hydroxide solution on haemoglobin film surface again, generate the immobilized haemoglobin biology sensor of silicon sol-gel film through the room temperature base catalyzed polymerization.
2,, it is characterized in that silane is selected from tetraethoxysilane or tetramethoxy-silicane as the said method of claim 1.
3,, it is characterized in that alcohol is selected from ethanol or methyl alcohol as the said method of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310117748 CN1635370A (en) | 2003-12-30 | 2003-12-30 | Method for preparing biosensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310117748 CN1635370A (en) | 2003-12-30 | 2003-12-30 | Method for preparing biosensor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1635370A true CN1635370A (en) | 2005-07-06 |
Family
ID=34843714
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200310117748 Pending CN1635370A (en) | 2003-12-30 | 2003-12-30 | Method for preparing biosensor |
Country Status (1)
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|---|---|
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104977342A (en) * | 2015-06-18 | 2015-10-14 | 青岛科技大学 | Graphene-exfoliation hydrotalcite-like compound composite material immobilized protein modified electrode, production method and application thereof |
| CN106706524A (en) * | 2017-02-22 | 2017-05-24 | 大连大学 | Method of quickly detecting oxygen content in gas by using fibroin membrane |
| CN110346433A (en) * | 2018-04-08 | 2019-10-18 | 海南师范大学 | A kind of preparation method and application of hemoglobin (Hb)/titanate radical nanopipe (TNTs) modified electrode |
-
2003
- 2003-12-30 CN CN 200310117748 patent/CN1635370A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104977342A (en) * | 2015-06-18 | 2015-10-14 | 青岛科技大学 | Graphene-exfoliation hydrotalcite-like compound composite material immobilized protein modified electrode, production method and application thereof |
| CN104977342B (en) * | 2015-06-18 | 2017-11-07 | 青岛科技大学 | Graphene peels off houghite piece composite ankyrin modified electrode and its preparation method and application |
| CN106706524A (en) * | 2017-02-22 | 2017-05-24 | 大连大学 | Method of quickly detecting oxygen content in gas by using fibroin membrane |
| CN106706524B (en) * | 2017-02-22 | 2019-04-19 | 大连大学 | A method of using oxygen content in the quick detection gas of fibroin film |
| CN110346433A (en) * | 2018-04-08 | 2019-10-18 | 海南师范大学 | A kind of preparation method and application of hemoglobin (Hb)/titanate radical nanopipe (TNTs) modified electrode |
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