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Antibodies against SARS-CoV-2 and uses thereof

A sars-cov-2 and antibody technology, applied in the direction of antibodies, applications, antiviral agents, etc., can solve problems such as limited clinical effects

Pending Publication Date: 2021-11-02
YANG SHENG TANG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patients with pneumonia caused by SARS-CoV-2 infection should only be given general supportive treatment, oxygen therapy measures and antiviral treatment, such as α-interferon, lopinavir / ritonavir, chloroquine phosphate, etc. These measures bring limited clinical efficacy

Method used

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  • Antibodies against SARS-CoV-2 and uses thereof
  • Antibodies against SARS-CoV-2 and uses thereof
  • Antibodies against SARS-CoV-2 and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0200] Example 1: Preparation of rabbit monoclonal antibody against SARS-CoV-2 receptor binding region RBD protein

[0201] 1.1 Preparation and activity identification of RBD protein in the receptor binding region of SARS-CoV-2:

[0202] The SARS-CoV-2 receptor binding region RBD-mFc was purchased from Beijing Yiqiao Shenzhou Technology Co., Ltd., and the RBD gene of the SARS-CoV-2 receptor binding region RBD-His protein was obtained from the NCBI database (GenBank ID: MN908947.3 ), with a His tag fused at the C-terminus. Referring to the full gene sequence of SARS-CoV2-2 (MN908947.3), the full-length sequence of the outer region of the SARS-CoV2-2 spike protein (corresponding to the viral Spike gene aa316-aa550) amino acid sequence was optimized according to human preferred codons to obtain its optimized code nucleic acid sequence. The N-terminal of the RBD nucleic acid sequence is connected to the signal peptide coding sequence, the C-terminal is connected to the optimized...

Embodiment 2

[0223] Example 2: Reactivity of anti-SARS-CoV-2 receptor binding domain protein RBD rabbit monoclonal antibody with SARS-CoV-2 receptor binding domain protein RBD

[0224] 2.1 Preparation of reaction plate

[0225] The SARS-CoV2-RBD (mFc tag) protein was treated with 50mM CB buffer (NaHCO 3 / Na 2 CO 3 Buffer, the final concentration is 50mM, the pH value is 9.6), and the final concentration is 2μg / mL; add 100μL of coating solution to each well of a 96-well microplate plate, and coat at 2~8℃ for 16~24 hours before Coat at 37°C for 2 hours; wash once with PBST washing solution (20mM PB7.4, 150mMNaCl, 0.1% Tween20); then add 200μL of blocking solution (pH containing 20% ​​calf serum and 1% casein) to each well 20mM Na at 7.4 2 HPO 4 / NaH 2 PO 4 buffer solution), placed at 37°C for blocking for 2 hours; discard the blocking solution. After drying, put it into an aluminum foil bag and store it at 2-8°C for later use.

[0226] 2.2 ELISA detection of RBD rabbit monoclonal an...

Embodiment 3

[0229] Example 3: Anti-SARS-CoV-2 receptor binding region protein RBD rabbit monoclonal antibody and SARS-CoV-2 receptor binding region protein RBD affinity constant detection

[0230] Kinetic analysis of the binding of monoclonal antibody and antigen using Biacore 8K system, all steps are carried out under PBS buffer, using the company's matching Protein A chip to capture monoclonal antibody diluted to 5 μg / mL, using SARS-CoV -2 Receptor binding region protein RBD-His is used as the detection antigen, and the antigen is diluted into five gradients of 100nM, 50nM, 25nM, 12.5nM, and 6.75nM respectively during detection. The detection is carried out according to the following procedure: capture (Captue) for 60s, analyze ( Analyte) 300s, dissociation (Dissociation) 600s, regeneration (Regeneration) 60s. The data acquisition and analysis software supporting the instrument was used to calculate the affinity equilibrium dissociation constant.

[0231] The result is as Figure 6 Sh...

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Abstract

The invention relates to the fields of immunology and molecular virology, in particular to the fields of diagnosis, prevention and treatment of SARS-CoV-2. In particular, the present invention relates to monoclonal antibodies against SARS-CoV-2, as well as compositions (e.g., diagnostic agents and therapeutic agents) comprising the antibodies. In addition, the invention also relates to application of the antibody. The antibodies of the invention can be used for diagnosing, preventing and / or treating SARS-CoV-2 infection and / or diseases caused by the infection (e.g., COVID-19).

Description

technical field [0001] The present invention relates to the field of immunology and molecular virology, especially the field of diagnosis, prevention and treatment of SARS-CoV-2. In particular, the invention relates to anti-SARS-CoV-2 monoclonal antibodies, and compositions (eg, diagnostic and therapeutic agents) comprising such antibodies. Furthermore, the present invention also relates to the use of said antibody. Antibodies of the invention can be used to diagnose, prevent and / or treat SARS-CoV-2 infection and / or diseases caused by said infection (eg, COVID-19). Background technique [0002] Coronavirus (coronavirus) infection can lead to respiratory diseases in humans. Mild coronavirus infection can cause flu-like symptoms, and severe infection can develop into severe viral pneumonia, threatening human life and health. Coronaviruses can infect humans and animals at the same time. If some animal-derived coronaviruses break through the host barrier and infect humans, the...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13G01N33/569G01N33/577A61K39/42A61P31/14
CPCC07K16/10G01N33/56983G01N33/577A61P31/14C07K2317/565C07K2317/56C07K2317/24C07K2317/567C07K2317/92C07K2317/52C07K2317/76G01N2333/165G01N2469/10A61K2039/505
Inventor 陈毅歆罗文新洪俊平陈俊煜蒋艺超张雅丽熊华龙吴倩王倩张天英袁权夏宁邵
Owner YANG SHENG TANG
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