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An electrochemical immunosensor for detecting cp4-epsps protein and its preparation method and application

A CP4-EPSPS, immunosensor technology, applied in the field of electrochemical immunosensor and its preparation, to achieve reliable electrochemical activity, high sensitivity, light and easy to carry the effect of the instrument

Active Publication Date: 2019-05-31
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no technology to use nanobodies to construct electrochemical immunosensors to achieve efficient detection of CP4-EPSPS protein.

Method used

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  • An electrochemical immunosensor for detecting cp4-epsps protein and its preparation method and application
  • An electrochemical immunosensor for detecting cp4-epsps protein and its preparation method and application
  • An electrochemical immunosensor for detecting cp4-epsps protein and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation of AuNPs-CMK-3

[0048] (1) Preparation of AuNPs

[0049] Take 100mL of HAuCl with a concentration of 0.12mg / mL 4 The solution was heated and boiled in a condensing reflux device, and 2.5mL of 38.8mM C 6 h 5 Na 3 o 7 .2H 2 O solution was used as a reducing agent. After stirring and heating for 15 min, the color changed to orange-red, and the heating source was removed and gradually cooled to room temperature to obtain the AuNPs solution.

[0050] (2) Preparation of AuNPs-CMK-3

[0051] Weigh 30mg of CMK-3 and add it to the above 100mL AuNPs solution, fully stir the reaction at room temperature for 4h, and collect the suspension by vacuum filtration through a filter membrane with a pore size of 0.22μm. Dry for 1h. The above dried AuNPs-CMK-3 was weighed and dissolved in DMF to prepare an AuNPs-CMK-3 dispersion with a final concentration of 0.5 mg / mL.

Embodiment 2

[0052] Example 2 Preparation of AuNPs-CMK-3

[0053] (1) Preparation of AuNPs

[0054] Take 50mL of HAuCl with a concentration of 0.12mg / mL 4 The solution was heated and boiled in a condensing reflux device, and 1.25mL of 38.8mM C 6 h 5 Na 3 o 7 .2H 2 O solution was used as the reducing agent, and after stirring and heating for 10 min, the heating source was removed and gradually cooled to room temperature to obtain the AuNPs solution.

[0055] (2) Preparation of AuNPs-CMK-3

[0056] Weigh 30mg of CMK-3 and add it to the above 50mL AuNPs solution, fully stir the reaction at room temperature for 4h, and collect the suspension by vacuum filtration through a filter membrane with a pore size of 0.22μm. Dry for 1h. The above dried AuNPs-CMK-3 was weighed and dissolved in DMF to prepare an AuNPs-CMK-3 dispersion with a final concentration of 0.5 mg / mL.

Embodiment 3

[0057] Example 3 Preparation of AuNPs-CMK-3

[0058] (1) Preparation of AuNPs

[0059] Take 200mL of HAuCl with a concentration of 0.12mg / mL 4 The solution was heated and boiled in a condensing reflux device, and 5mL of 38.8mM C 6 h 5 Na 3 o 7 .2H 2 O solution was used as a reducing agent, and after stirring and heating for 30 min, the heating source was removed and gradually cooled to room temperature to obtain the AuNPs solution.

[0060] (2) Preparation of AuNPs-CMK-3

[0061] Weigh 30mg of CMK-3 and add it to the above 200mL AuNPs solution, fully stir the reaction at room temperature for 4h, and collect the suspension by vacuum filtration through a filter membrane with a pore size of 0.22μm. Dry for 1h. The above dried AuNPs-CMK-3 was weighed and dissolved in DMF to prepare an AuNPs-CMK-3 dispersion with a final concentration of 0.5 mg / mL.

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Abstract

The invention discloses an electrochemical immunosensor for detecting a CP4-EPSPS protein, and a preparation method and an application thereof. The surface of a substrate electrode is modified with a AuNPs-CMK-3 dispersion, then is activated with an EDC and NHS mixed solution, and finally is covalently bonded with a CP4-EPSPS antibody and thionine mixed solution. The substrate element is a glassy carbon electrode, and the CP4-EPSPS antibody is a nanometer antibody. The electrochemical immunosensor has the obvious advantages of high sensitivity, high specificity, and portability in device, plays a great role in the detection of bio-macromolecules, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of rapid detection of CP4-EPSPS protein, and in particular relates to an electrochemical immunosensor for detecting CP4-EPSPS protein and its preparation method and application. Background technique [0002] EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) is a key enzyme in the shikimate pathway, which widely exists in plants and microorganisms, and is the target enzyme of glyphosate, a broad-spectrum high-efficiency herbicide. CP4-EPSPS has very high resistance to glyphosate, so it is widely used in crop transgenic engineering. Since the commercialization of the first transgenic glyphosate-resistant soybean 40-3-2 in 1996, the acreage of glyphosate-resistant crops has grown rapidly. Facing a large number of transgenic crops containing CP4-EPSPS, we urgently need to establish a scientific and sensitive detection method for qualitative and quantitative detection. [0003] Currently, methods for detectin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/26
CPCG01N27/26G01N27/3278
Inventor 沈艳飞张明明潘登陈慧琴张袁健
Owner SOUTHEAST UNIV