Novel coronavirus neutralizing antibody magnetic bead fluorescence detection kit and detection method

A coronavirus and fluorescence detection technology, which is applied in the direction of virus/bacteriophage, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of high detection cost, insufficient sensitivity, and no cost-effective advantages, so as to achieve good detection accuracy Sexuality and the effect of reducing the cost of testing

Active Publication Date: 2021-12-14
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the lack of sensitivity of the current detection method for serum neutralizing antibodies of the new coronavirus and the large demand for recombinant proteins in the current functional ELISA detection method, resulting in high detection costs and no cost-effective advantages, the present invention provides a new type of coronavirus. And antibody magnetic bead fluorescence detection kit and detection method thereof

Method used

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  • Novel coronavirus neutralizing antibody magnetic bead fluorescence detection kit and detection method
  • Novel coronavirus neutralizing antibody magnetic bead fluorescence detection kit and detection method
  • Novel coronavirus neutralizing antibody magnetic bead fluorescence detection kit and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0060] The critical concentration at which the RBD-mFc protein binds to most of the ACE2 protein magnetic beads at a high level The specific operation steps are as follows:

[0061] (1) Dilute the RBD-mFc recombinant protein into a series of concentration gradients with the sample diluent, the concentrations are 2ng / mL, 4ng / mL, 8ng / mL, 12ng / mL and 16ng / mL. Dilute the mouse IgG1 isotype control antibody to a concentration of 16 ng / mL with sample diluent.

[0062] (2) Add RBD-mFc and mouse IgG isotype control antibody at various concentrations diluted in step (1) to a round-bottom 96-well plate at 50 μL / well, and conduct experiments in duplicate wells for each concentration sample.

[0063] (3) Resuspend the ACE2 magnetic beads, absorb the ACE2 magnetic bead solution at 1 μL / reaction, wash the ACE2 magnetic beads in a round-bottom 96-well plate with 200 μL of sample diluent, and wash twice. When the cleaning solution is sucked and discarded, the magnetic beads are retained in t...

Embodiment 2

[0070] The influence of different concentrations of RBD-mFc on the sensitivity of the kit of the present invention to detect the neutralizing antibody of the new coronavirus

[0071] The program operation steps of this embodiment are as follows:

[0072] (1) Use the sample diluent to dilute the positive standard and negative standard of the new coronavirus neutralizing antibody respectively to a concentration of 5ng / mL, 15ng / mL, 45ng / mL, 135ng / mL, 400ng / mL, 1200ng / mL and For 3600ng / mL antibody diluent, add 50 μL / well of the positive standard diluent and negative standard diluent of each concentration of the new coronavirus neutralizing antibody to the round-bottom 96-well plate, and add duplicate wells for each antibody concentration;

[0073] (2) Dilute RBD-mFc and mouse IgG isotype control antibodies with sample diluent to concentrations of 4 ng / mL, 100 ng / mL, 200 ng / mL and 2000 ng / mL respectively, and dilute each concentration of RBD-mFc and mouse IgG isotype Add 50 μL / wel...

Embodiment 3

[0083] Sensitivity evaluation of ACE2 magnetic beads / RBD-mFc novel coronavirus neutralizing antibody detection kit

[0084] In this example, a commercially available novel coronavirus neutralizing antibody function ELISA detection kit (SARS-CoV-2 Surrogate Virus Neutralization Test Kit product number: L00847-A produced by GenScript Biotechnology Co., Ltd. Item No.: L00847A) is used as a reference to evaluate the detection sensitivity of the ACE2 magnetic beads / RBD-mFc new coronavirus neutralizing antibody detection kit established by the present invention. In this example, a humanized monoclonal antibody (Acrobiosystems, Cat. No.: SAD-S35) with neutralizing activity of the new coronavirus and a human IgG1 isotype control antibody "Human IgG 1, K Isotype Control" ( BioLegend, Cat. No. Cat403502) as a positive standard for neutralizing antibodies to the new coronavirus and neutralizing antibodies to the new coronavirus in the ACE2 magnetic beads / RBD-mFc neutralizing antibody det...

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Abstract

The invention relates to the technical field of immunoassay detection, and discloses a novel coronavirus neutralizing antibody magnetic bead fluorescence detection kit and detection method. The kit comprises ACE2 protein magnetic beads, novel coronavirus S1 protein RBD structural domain-mouse IgG Fc fragment fusion protein, a fluorescent substance labelled animal anti-mouse IgG antibody, a novel coronavirus neutralizing antibody positive standard substance, a novel coronavirus neutralizing antibody negative standard substance, a mouse IgG homotype control antibody, a 10x concentrated washing solution and a sample diluent. The detection method implemented by the kit has relatively high sensitivity on the detection of the novel coronavirus neutralizing antibody. The kit is low in cost and high in cost performance when used for detecting the novel coronavirus neutralizing antibody, and the kit can fill the gap of high-sensitivity products in the current rapid detection kit market for the novel coronavirus neutralizing antibody.

Description

technical field [0001] The invention relates to the technical field of immune analysis and detection, in particular to a novel coronavirus neutralizing antibody magnetic bead fluorescence detection kit and a detection method thereof. Background technique [0002] New Coronary Pneumonia (COVID-19) is an infectious disease caused by a novel coronavirus (SARS-COV-2) infection. The disease is highly contagious, and the virus spreads and mutates quickly. As of July 16, 2021, the cumulative number of cases of new coronary pneumonia infection worldwide has reached 189 million, and the death toll due to infection has reached 4.08 million. COVID-19 has become a major infectious disease that seriously threatens human health. [0003] At present, there is no effective virus-specific chemical drug for the clinical treatment of new coronary pneumonia. A large number of clinical and real-world research results show that vaccines are the most effective weapon for humans to fight against...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/573G01N33/58G01N33/543G01N15/14C07K19/00
CPCG01N33/56983G01N33/573G01N33/582G01N33/54326G01N15/14C07K14/005G01N2333/165G01N2469/20G01N2015/1486C12N2770/20022C07K2319/30Y02A50/30
Inventor 孟庆来要翔宇吴长新傅羽佳
Owner SHANXI UNIV
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