Preparation method and application of electrochemical immunosensor for detecting 17 beta-estradiol

An immunosensor, estradiol technology, applied in the direction of material electrochemical variables, biological testing, material inspection products, etc., can solve the problems of complex instrument operation, complex sample processing, and high detection cost, and achieve simple operation, high sensitivity, and electrical conductivity. powerful effect

Inactive Publication Date: 2014-03-26
UNIV OF JINAN
View PDF3 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the methods for detecting 17β-estradiol mainly include high performance liquid chromatography, gas chromatography, etc., but the

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for preparing an electrochemical immunosensor for detecting 17β-estradiol, the steps are as follows:

[0026] (1) Dissolve 5 mg GO-PANI in 5 mL DMF by ultrasonication for 1 h, add 20 μL of GO-PANI solution dropwise on the surface of the glassy carbon electrode, and dry at room temperature;

[0027] (2) Dip the modified electrode in 1 mM HAuCl 4 and 1 mM H 2 PtCl 6 0.2 M Na 2 SO 4 In the solution, the scanning reduction was carried out by the constant potential method, the scanning potential was -0.2 V, and the scanning time was 300 s.

[0028] (3) 10 μl, 10 μg / mL 17β-estradiol complete antigen was added dropwise on the surface of the modified electrode, and dried at room temperature for 5 h.

[0029] (4) Residual active sites of basal planes blocked with 10 μl BSA (1%). After each step of modification, curve the electrode with PBS buffer solution to obtain the prepared immunosensor electrode.

[0030] (5) Add 10 μl of antibody-labeled layer and 10 μl o...

Embodiment 2

[0033] (1) Dissolve 5 mg GO-PANI in 5 mL DMF by ultrasonication for 1 h, add 20 μL of GO-PANI solution dropwise on the surface of the glassy carbon electrode, and dry at room temperature;

[0034] (2) Dip the modified electrode in 1 mM HAuCl 4 and 1 mM H 2 PtCl 6 0.2 M Na 2 SO 4 In the solution, the scanning reduction is carried out by the constant potential method, the scanning potential is -0.2 V, and the scanning time is 200 s;

[0035] (3) Add 10 μl, 10 μg / mL 17β-estradiol complete antigen dropwise on the modified electrode surface, and dry at room temperature for 5 h;

[0036] (4) Residual active sites of basal planes blocked with 10 μl BSA (1%). After each step of modification, use PBS buffer solution to curve the electrode to obtain the prepared immunosensor electrode;

[0037] (5) Add 10 μl of antibody-labeled layer and 10 μl of free 17β-estradiol standard on the prepared electrode surface, incubate at room temperature for 30 min, and wash unbound antibody and s...

Embodiment 3

[0040] (1) Dissolve 5 mg GO-PANI in 5 mL DMF by ultrasonication for 1 h, add 20 μL of GO-PANI solution dropwise on the surface of the glassy carbon electrode, and dry at room temperature;

[0041] (2) Dip the modified electrode in 1 mM HAuCl 4 and 1 mM H 2 PtCl 6 0.2 M Na 2 SO 4 In the solution, the scanning reduction is carried out by the constant potential method, the scanning potential is -0.2 V, and the scanning time is 200 s;

[0042] (3) Add 10 μl, 10 μg / mL 17β-estradiol complete antigen dropwise on the modified electrode surface, and dry at room temperature for 5 h;

[0043] (4) Residual active sites of basal planes blocked with 10 μl BSA (1%). After each step of modification, use PBS buffer solution to curve the electrode to obtain the prepared immunosensor electrode;

[0044] (5) Add 10 μl of antibody-labeled layer and 10 μl of free 17β-estradiol standard on the prepared electrode surface, incubate at room temperature for 30 min, and wash unbound antibody and s...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides an electrochemical immunosensor for detecting 17 beta-estradiol. According to the immunosensor, an electrode surface is modified by a prepared graphene oxide-polyaniline (GO-PANI) compound through an electrode surface modification technology, a platinum-gold alloy (Au-PtNPs) layer is polymerized on the electrode surface with an electro-polymerization method, 17 beta-estradiol complete antigen is modified on the electrode surface, and an antibody compound (PDA-PtNPs-AuNPs-Ab-HRP) and a 17 beta-estradiol standard substance are simultaneously modified on the electrode surface, so that the complete antigen and the standard substance simultaneously compete and combine with an antibody on an antibody compound, a signal is generated by an HRP catalytic substrate H2O2 combined on the antibody compound on the electrode surface, a working curve is drawn according to the relation between signal changes before and after adding of the substrate H2O2 and standard substance concentration, and the 17 beta-estradiol is detected. A PDA-PtNPs-AuNPs nano material is synthesized for the first time, the conductivity of the GO-PANI compound is high, and the Au-PtNPs nano alloy has excellent biocompatibility and certain catalytic performance on H2O2; and the electrochemical immunosensor has the advantages of high sensitivity, high detection speed and short detection period, and an actual sample can be detected rapidly and conveniently.

Description

technical field [0001] The invention belongs to the field of detection of nanometer materials and food additives, and provides an electrochemical immunosensor for detecting 17β-estradiol, which can be used for detecting 17β-estradiol in food. Background technique [0002] Estrogen is a G 18 steroid hormone. There are three types of estrogen found in the human body: estradiol, estrone and estriol, among which estradiol has the strongest biological activity. Estradiol can bind to estrogen receptors in the body and change the normal physiological functions of the endocrine system. Because of its anabolic effect, it is used in livestock production to promote growth, while low concentrations of estrogen can cause abnormalities in human sexual development and reduce the number of human sperm. In some countries of the European Union, estrogen has been banned for use in animal husbandry. Therefore, it is urgent to establish a rapid and convenient detection method to detect the con...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N27/30G01N33/53
Inventor 黄加栋李杰刘素
Owner UNIV OF JINAN
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap