Method for maintaining antigen activity in antigen-fluorescent microsphere conjugate and application in novel coronavirus antibody chromatography detection reagent

A fluorescent microsphere and detection reagent technology, applied in the field of immunochromatography, can solve the problems of decreased antigen activity, low sensitivity of antibody detection reagents, inability to stretch protein conformation, etc.

Active Publication Date: 2020-10-23
SICHUAN XINCHENG BIOLOGICAL CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is: in the coupling of the existing antigen, after the antigen is coupled to the surface of the microsphere, it may also be due to steric hindrance, so that the protein conformation cannot b

Method used

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  • Method for maintaining antigen activity in antigen-fluorescent microsphere conjugate and application in novel coronavirus antibody chromatography detection reagent
  • Method for maintaining antigen activity in antigen-fluorescent microsphere conjugate and application in novel coronavirus antibody chromatography detection reagent
  • Method for maintaining antigen activity in antigen-fluorescent microsphere conjugate and application in novel coronavirus antibody chromatography detection reagent

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Experimental program
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Embodiment 1

[0038] 1. Preparation of antigen-antigen-fluorescent microsphere conjugates

[0039](1) Take out 500ul fluorescent microspheres (1% concentration W / V, green fluorescence-excitation 475nm-emission 525nM) into a centrifuge tube.

[0040] (2) Centrifuge (12000rpm-20000rpm according to different particle sizes) for 10min to settle the microspheres and remove the supernatant.

[0041] (3) Add 500ul coupling buffer (50mM MES pH 6.0) and mix well.

[0042] (4) Add 20ul EDC solution (200mM), 20ul sulfo-NHS solution (200mM), mix well, and incubate on a rotary mixer for 30min.

[0043] (5) Centrifuge (12000rpm-20000rpm according to different particle sizes) for 10min to settle the microspheres and remove the supernatant.

[0044] (6) Add 500ul coupling buffer (50mM MES ph6.0), mix well, add 0.1mg new crown antigen, mix well, and incubate for 1 hour at room temperature with a rotary mixer.

[0045] (7) Centrifugation (choose 12000rpm-20000rpm according to different particle sizes

[...

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Abstract

The invention discloses a method for maintaining antigen activity in an antigen-fluorescent microsphere conjugate and application in a novel coronavirus antibody chromatography detection reagent, andsolves the problem that in the existing antigen coupling, after the antigen is coupled to the surface of the microsphere, the protein conformation cannot extend possibly because of steric hindrance and thus the antigen activity is reduced, a part with immunocompetence in an antigen structure is extremely likely to participate in a coupling chemical reaction to cause reduction of the antigen activity and thus the sensitivity of the antibody detection reagent is low. The method comprises the following steps: coupling fluorescent microspheres with an antigen to obtain an antigen-fluorescent microsphere conjugate, and then coupling the antigen again to obtain the antigen-antigen-fluorescent microsphere conjugate. The method has the advantages of high antigen activity of novel coronavirus, sensitive detection, high accuracy and the like.

Description

technical field [0001] The invention relates to the technical field of immunochromatography, in particular to a method for maintaining antigen activity in an antigen-fluorescent microsphere conjugate and its application in a novel coronavirus antibody chromatography detection reagent. Background technique [0002] At present, the detection methods for the new coronavirus are mainly divided into two categories, one is the direct detection of the virus, such as the "gold standard" nucleic acid detection of etiology, which is the detection of certain specific nucleic acid sequences in the virus gene; the other is the indirect detection of the virus. Detection, such as the detection of new coronavirus-specific serum antibodies, when the virus invades the human body, the human immune system will produce a large number of immunoglobulins, the most common of which are IgG and IgM, which are key components of the body's immune response to the virus, so through Serum testing for susp...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/569G01N33/558G01N33/533
CPCG01N33/533G01N33/558G01N33/56983G01N33/577G01N33/582G01N33/587G01N33/6854G01N2333/165G01N2469/20
Inventor 林源蒋明君李学锐
Owner SICHUAN XINCHENG BIOLOGICAL CO LTD
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