Bispecific antibody for resisting novel coronavirus and application of bispecific antibody

A bispecific antibody, coronavirus technology, applied in the fields of biotechnology and immunology, can solve problems such as application limitations, and achieve the effect of improving selectivity and neutralizing activity, reducing difficulty and good stability

Active Publication Date: 2021-10-29
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hybrid-hybridoma method is to produce bispecific monoclonal antibodies by means of cell hybridization or ternary hybridomas. These cell hybridomas or ternary hybridomas are fused by established hybridomas, or established hybridomas and hybridomas derived from mice The fusion of lymphocytes can only be used to produce murine bispecific antibodies, so its application is greatly limited

Method used

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  • Bispecific antibody for resisting novel coronavirus and application of bispecific antibody
  • Bispecific antibody for resisting novel coronavirus and application of bispecific antibody
  • Bispecific antibody for resisting novel coronavirus and application of bispecific antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1 Bispecific antibody construction

[0080] In the construction of bispecific antibodies, the arrangement of the variable regions of the two antibodies will affect their ability to bind to antigens, and different constructions may lead to differences in antiviral activity. Therefore, whether the bispecific antibody constructed based on antibodies at two different sites can retain its dual-site binding activity and whether the antiviral activity can be improved still has great uncertainty. Bispecific antibodies are fusion proteins formed between antibodies with different structures. Antibodies with different structures bind to different antigens of two cells or different sites of the same antigen to exert different functions. In order to enable each antibody in the bispecific antibody to fully exert its respective functions, a large number of candidate bispecific antibodies were designed in this example in order to minimize the impact of the sequence and structur...

Embodiment 2

[0095] Example 2 Construction of Recombinant Expression of Bispecific Antibody and Purification of Protein Expression

[0096] In this example, two different bispecific antibodies, BS-mAb-1 and BS-mAb-2, were constructed based on the two novel coronavirus S antigen-specific antibodies H4 and B38, and their heavy chain amino acid sequences As shown in SEQ ID NO.2 and SEQ ID NO.6, its light chain amino acid sequence is shown in SEQ ID NO.4 and SEQ ID NO.8 respectively.

[0097] HEK293T cells (ATCC CRL-3216) were cultured in DMEM containing 10% FBS. HEK 293T cells were co-transfected with the above-mentioned recombinant expression vectors of BS-mAb-1 and BS-mAb-2, which respectively encode the heavy chain and light chain of the antibody. After 4-6 hours of transfection, the cell culture medium was replaced with serum-free DMEM, and the culture was continued for 3 days. The supernatant was collected, then supplemented with DMEM, continued to culture for 4 days, and then the supe...

Embodiment 3

[0099] Example 3 Evaluation of the two-site binding ability of BS-mAb-1 and BS-mAb-2 antibodies to the S protein RBD

[0100]In this example, the ForteBio Octet RED 96 biofilm layer surface interference technology was used to analyze the binding of the purified antibody to the S protein RBD, and to evaluate whether the BS-mAb-1 and BS-mAb-2 antibodies have two-site binding ability .

[0101] The S protein RBD (Science. 2020 Jun 12; 368(6496): 1274-1278) with a histidine tag was immobilized on the probe (Fortebio). In order to evaluate the two-site binding ability of BS-mAb-1 antibody, the probe immobilizing RBD protein was first combined with H4 antibody (Science. 2020 Jun 12; 368(6496):1274-1278) and allowed to saturate. Afterwards, BS-mAb-1 and equal concentration of H4 antibody mixture were passed through the H4 antibody to saturate the bound probe. The results showed that compared with the binding level of the H4-H4 antibody to the RBD, the RBD saturated with the H4 anti...

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Abstract

The invention relates to a bispecific antibody for resisting novel coronavirus and application of the bispecific antibody. The bispecific antibody is obtained by modifying new coronavirus monoclonal antibodies H4 and B38 through a genetic engineering method, different sites of novel coronavirus S protein RBD can be recognized at the same time, the neutralizing activity on new coronavirus pseudovirus is far higher than that of a maternal monoclonal antibody, and the inhibitory activity on new coronavirus live virus is further higher than that of the maternal monoclonal antibody. According to the bispecific antibody for resisting novel coronavirus, the selectivity and neutralizing activity of the maternal monoclonal antibody are improved, and the safety and effectiveness of monoclonal antibody drugs are improved; and the bispecific antibody can be used for preparing potential drugs for diagnosing, preventing and treating diseases caused by novel coronavirus and is huge in market value and good in application prospect.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and immunology, in particular to an anti-new coronavirus bispecific antibody and its preparation method and application. Background technique [0002] The spike protein (Spike, S protein) on the surface of the new coronavirus (2019-nCoV) binds to the host cell receptor angiotensin-converting enzyme 2 (ACE2) molecule during the infection of the host, thereby initiating the fusion of the viral membrane and the host cell membrane , causing the host cell to be infected with the virus. The S protein is divided into two parts, S1 and S2. Studies have confirmed that the receptor binding domain (RBD) of the C-terminal (CTD) of S1 binds to ACE2 and mediates the process of membrane fusion. [0003] So far, neutralizing antibodies have proven to be effective treatments for viral diseases. Drugs currently on the market for the treatment and prevention of viral infections include palivizumab (Synagis)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/46C12N15/13C12N15/85A61K39/42A61P31/14
CPCC07K16/10C12N15/85A61P31/14C07K2317/31C07K2317/76C07K2317/565C07K2317/622C07K2317/94C12N2800/107C12N2800/22A61K2039/505
Inventor 高福吴燕李世华黎朝晖谭曙光
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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