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Electrochemical enzyme-linked immunosensor, preparation thereof and application for detecting antigens

An enzyme-linked immunosorbent and electrochemical technology, applied in the field of protein detection, can solve the problems of low specificity, low detection sensitivity and accuracy of electrochemical immunosensors, and achieve high sensitivity, good accuracy, and low detection limit Effect

Active Publication Date: 2019-10-25
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] The invention solves the technical problems of low detection sensitivity and accuracy and low specificity of the electrochemical immunosensor in the prior art

Method used

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  • Electrochemical enzyme-linked immunosensor, preparation thereof and application for detecting antigens
  • Electrochemical enzyme-linked immunosensor, preparation thereof and application for detecting antigens
  • Electrochemical enzyme-linked immunosensor, preparation thereof and application for detecting antigens

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Embodiment 1

[0053] The preparation of the electrochemical enzyme-linked immunosensor and the method for the electrochemical immunological detection of the antigen by highly sensitive alkaline phosphatase-labeled magnetic beads in the present invention have the following processes and steps:

[0054] (1) Preparation of ZnO@rGO / CPE

[0055] Based on the improved Kovtyukhova two-step oxidation exfoliation method, graphene oxide gel (GO, 1.0-100mg mL -1 ), take 0.1-5.0mL GO and ultrasonically disperse it in 9.0mL of water, then add 10-80mg Zn powder and 0.1-5.0mL NH 4 Cl aqueous solution (0.2g mL -1 ), magnetically stirred at an appropriate speed for 1-60min, then left to stand, continued to react for 30min, then centrifuged and washed the black product with ultrapure water for at least 6 times, and finally dried the final product (ZnO@ rGO).

[0056] Accurately weigh 0.05g ZnO@rGO and 0.95g graphite powder and mix them evenly in the grinding platinum, add 200μL paraffin oil and mechanical...

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Abstract

The invention relates to an electrochemical enzyme-linked immunosensor, a preparation thereof and an application for detecting antigens, which belong to the technical field of protein detection. According to the preparation, an electrochemical enzyme-linked immunoprobe is prepared, the antigens are added into a biological conjugate of a magnetic bead and captured antibodies for incubation, an upper clear liquid is removed under the condition of an externally applied magnetic field, then the antigens are added into a biological conjugate of nano gold, detection antibodies and enzyme for incubation, and the electrochemical enzyme-linked immunoprobe is obtained after the double-antibody sandwich reaction; and a dispersion liquid of the immunoprobe is modified on the surface of an electrode ofa carbon paste base, so as to obtain the electrochemical enzyme-linked immunosensor. A linearity range of the electrochemical enzyme-linked immunosensor (MB-eElisa) provided by the invention is 0.01-6.0 ng mL<-1>, the detection limit is 4.0 pg mL<-1>(S / N=3); and compared with the existing electrochemical immunodetection method, the preparation method has low detection limit and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of protein detection, and more specifically relates to an electrochemical enzyme-linked immunosensor and its preparation and application of antigen detection. Background technique [0002] Antigen refers to a class of substances that can stimulate the immune system of living organisms, induce immune feedback, produce antibodies for humoral immunity or effector lymphocytes for cellular immunity, and react with them in vivo and in vitro. In addition, antigens have very important theoretical research significance in the diagnosis and clinical treatment of tumor diseases. At present, there are many kinds of antigens related to tumor diseases, such as carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), prostate specific protein (PSA), and sugar chain antigen (CA19-9). The most widely applicable detection object for diseases, it is a cell surface glycoprotein with a molecular weight of about 200kDa, which is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N27/26
CPCG01N27/26G01N33/577
Inventor 吴康兵郝俊兴
Owner HUAZHONG UNIV OF SCI & TECH
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