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Novel coronavirus Delta mutant strain specific antibody and application thereof

A coronavirus and antibody technology, applied in the direction of virus antigen components, antibodies, antiviral agents, etc., can solve the problem of incapable of vaccine-specific detection and identification, and achieve the effect of accelerating the research and development process and high affinity

Active Publication Date: 2022-04-15
百斯医学诊断科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antibodies currently tested for the new coronavirus vaccine are all general-purpose antibodies, and the vaccines designed for the new coronavirus Delta mutant strain cannot be specifically detected and identified

Method used

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  • Novel coronavirus Delta mutant strain specific antibody and application thereof
  • Novel coronavirus Delta mutant strain specific antibody and application thereof
  • Novel coronavirus Delta mutant strain specific antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Preparation of Specific Novel Coronavirus Delta Antibody

[0073] In this example, the novel coronavirus Delta mutant strain-specific antibody was prepared according to the following method:

[0074] 1. Mice immunization: Spike protein of the new coronavirus Delta mutant strain (purchased from Acrobiosystems) was used as the immunogen, and mice were immunized with the new coronavirus Delta mutant strain Spike protein. After the immunization, the sera of the immunized animals were detected by ELISA method to determine the level of immune response. After routine immunization, if the immunized animal can reach the level of immune response against the immunogen, cell fusion is performed.

[0075] 2. Screening: Screen the supernatant of fusion cells by ELISA method, and select cells that are positive for the specific binding of Spike protein of the new coronavirus Delta mutant strain and have no binding with wild-type, Alpha, Beta, Gamma, and Omicron Spike protein...

Embodiment 2

[0080] Example 2 Antibody specificity analysis of novel coronavirus Delta mutant strain

[0081] In this example, 10 monoclonal antibodies with different sequences were obtained according to the method in Example 1 above, and the enzyme-linked immunosorbent assay was used to analyze the specificity of the above-mentioned novel coronavirus Delta mutant antibody. The analysis method is as follows:

[0082] 1. Dilute the S protein of the wild-type, Alpha, Beta, Gamma, and Delta of the new coronavirus with PBS to 2 μg / mL, and add it to the wells of the microplate, 100 μL per well. Seal the plate with a sealing film and place it at 4°C overnight.

[0083] 2. Discard the liquid in the well, pat dry the ELISA plate, wash the plate with PBST washing solution, soak in 300 μL / well, pat dry the ELISA plate, and perform the next wash, wash the plate 3 times in total.

[0084] 3. Add 100 μL of blocking agent (PBST washing solution containing 5% BSA) to each well, seal the plate with a sea...

Embodiment 3

[0094] Example 3 Analysis, identification and functional analysis of novel coronavirus Delta mutant strain-specific antibodies

[0095] In this example, the novel coronavirus Delta mutant strain-specific antibody (No. 3) screened in Example 2 was analyzed, identified and functionally analyzed using methods known in the art, as follows:

[0096] 1. SDS-PAGE identification results ( figure 1 ) shows that the reduction electrophoresis of antibody No. 3 has two bands with molecular weights of about 27kDa and 50kDa respectively, and the purity is greater than 99%.

[0097] 2. SEC-MALS identification results ( figure 2 ) shows that the No. 3 antibody has a purity greater than 99% and a molecular weight of 154kDa.

[0098] 3. ELISA binding experiment results ( image 3 ) shows that the antibody No. 3 can specifically recognize the 2019-nCoV Delta mutant antigen (Spike protein of the 2019-nCoV Delta mutant strain), but has no binding to wild-type, Alpha, Beta, Gamma, Omicron Spike...

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Abstract

The invention relates to the technical field of antibodies, in particular to a novel coronavirus Delta mutant strain specific antibody and application thereof. The invention provides an antibody specifically combined with a novel coronavirus Delta mutant strain, and the antibody can be used as a specific antibody for Delta mutant strain antigen detection, is used for identifying a novel coronavirus vaccine designed for the Delta mutant strain, and is used for quantitatively detecting the Spike protein content expressed by the vaccine. The antibody can be used for quality control of novel coronavirus vaccines and immunogenicity detection of clinical and preclinical research, or can be used as a quality control antibody for detection of protective antibodies in serum after vaccination.

Description

technical field [0001] The invention relates to the technical field of antibodies, in particular to novel coronavirus Delta mutant-specific antibodies and applications thereof. Background technique [0002] The 2019-nCoV Delta mutant is a highly transmissible 2019-nCoV mutant. Testing the protective power of existing vaccines in clinical research and designing highly specific vaccines based on mutated antigens are the key to dealing with the prevalence of novel coronavirus mutants. [0003] At present, many vaccine companies such as Pfizer, Moderna and AstraZeneca have conducted extensive research on novel coronavirus mutant strains to advance the development of a new generation of vaccines: Pfizer and BioNTech have carried out research on Delta and Alpha mutant strains, and is also testing a Beta-specific mRNA vaccine in a randomized, placebo-controlled clinical trial; Moderna is recruiting volunteers to test a multivalent vaccine with a Beta-Delta combination ; AstraZene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10G01N33/569A61K39/42A61P31/14
CPCC07K16/10G01N33/56983A61K39/12A61P31/14G01N2333/165C07K2317/56C07K2317/565A61K2039/505
Inventor 葛平菊苗景赟陈宜顶王恒玲焦秋伶任文林
Owner 百斯医学诊断科技(北京)有限公司
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