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Preparation method and application of electrochemical immunosensor based on HS-beta-CD-Ag-GOD conjugate

An immunosensor and conjugate technology, which is applied in the field of immunoassay and biosensing, can solve the problems of long detection period, many required reagents, complicated procedures, etc., and achieves improved detection sensitivity, good molecular recognition ability, and high conductivity. Effect

Active Publication Date: 2015-03-25
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to its long detection cycle, complex procedures and many reagents required, it is far from meeting the requirements of modern detection.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preparation of embodiment 1 HS-β-CD-Ag nanometer material

[0037] 1 mL, 0.005 mol / L of AgNO 3 and 1 mL, 0.02 mol / L trisodium citrate were added to 97 mL high-purity water, stirred evenly, and 1 mL, 1.5 mg / mL NaBH 4 , fully stirred for 20 min, added 0.0238 g of mercapto-β-cyclodextrin HS-β-CD, magnetically stirred for 24 h, centrifuged at 11,000 rpm for 10 min, and vacuum-dried at a constant temperature of 60 °C;

Embodiment 2

[0038] Example 2 Preparation of HS-β-CD-Ag nanomaterials

[0039] 1 mL, 0.01 mol / L of AgNO 3 and 1 mL, 0.03 mol / L trisodium citrate were added to 97 mL high-purity water, stirred evenly, and 1 mL, 1.79 mg / mL NaBH 4 , fully stirred for 20 min, added 0.0238 g of mercapto-β-cyclodextrin HS-β-CD, magnetically stirred for 24 h, centrifuged at 11,000 rpm for 10 min, and vacuum-dried at a constant temperature of 60 °C;

Embodiment 3

[0040] Example 3 Preparation of HS-β-CD-Ag nanomaterials

[0041] 1 mL, 0.015 mol / L of AgNO 3 and 1 mL, 0.04 mol / L trisodium citrate were added to 97 mL high-purity water, stirred evenly, and 1 mL, 2.0 mg / mL NaBH 4 , fully stirred for 20 min, added 0.0238 g of mercapto-β-cyclodextrin HS-β-CD, magnetically stirred for 24 h, centrifuged at 11,000 rpm for 10 min, and vacuum-dried at a constant temperature of 60 °C;

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Abstract

The invention belongs to the technical fields of immunoassay and biosensing and discloses a preparation method and application of an electrochemical immunosensor based on an HS-beta-CD-Ag-GOD conjugate. The electrochemical immunosensor is used for rapidly detecting a hog cholera virus antigen CSFV. The manufacturing scheme comprises the following steps: modifying a working electrode a bare platinum carbon electrode by using MWCNTs-CD-Fc-Ab1, and sequentially adding bovine serum albumin, hog cholera virus antigen CSFV and an Ab2-HS-beta-CD-Ag-GOD conjugate. The HS-beta-CD-Ag-GOD conjugate can convert glucose into gluconic acid, two protons and two electrons are transferred to oxygen molecules, hydrogen peroxide is generated, and an HS-beta-CD-Ag nanometer material serves as a mimic enzyme, the reduction of the hydrogen peroxide is catalyzed, the dual amplification of an electrochemical signal is realized, the high sensitivity is realized, and the detection limit can be low to 0.33pg / mL.

Description

technical field [0001] The invention belongs to the technical field of immune analysis and biosensing, in particular to the preparation and application of an electrochemical immunosensor based on HS-β-CD-Ag-GOD conjugates, and the development of a rapid detection of CSFV antigen Sandwich type electrochemical immunosensor. Background technique [0002] CSFV, or swine cholera, is an acute, febrile, highly contagious infectious disease characterized by high temperature and capillary degeneration that causes systemic hemorrhage, necrosis, and infarction. Due to its high pathogenicity and widespread mortality, it causes severe economic losses to the swine industry. Many countries have formulated veterinary regulations to list swine fever virus as one of the legal infectious diseases, that is, diseases that must be reported on the list. In the event of an epidemic or suspicious epidemic, it should be reported as soon as possible, and emergency veterinary health and epidemic prev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N27/327
CPCG01N27/3278G01N33/54366G01N33/54386G01N33/56983G01N2333/185
Inventor 吴丹高健魏琴闫涛马洪敏张勇罗川南杜斌庞雪辉
Owner UNIV OF JINAN
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