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Phage display antibody library and monoclonal antibody against new coronavirus sars-cov-2 obtained based on its panning

A monoclonal antibody, sars-cov-2 technology, applied in the fields of biomedicine and molecular biology, can solve problems such as toxic side effects, no obvious therapeutic effect, and difficult prevention, and achieve the effect of expanding powerful resources

Active Publication Date: 2021-08-10
山东宽和正生物医药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Compared with the original coronavirus, SARS-CoV-2 is not only contagious in patients with severe symptoms, but also in patients with mild symptoms and even in the incubation period. There are many ways of transmission, and it can be transmitted by droplets and contact. It is very difficult to prevent
[0003] Various countries have proposed a variety of treatment options for the treatment of new coronary pneumonia, mainly using the small molecule drug remdesivir, the combination of chloroquine and hydroxychloroquine, the combination of lopinavir and ritonavir, and the combination of lopinavir and ritonavir. Unfortunately, the three-drug combination of navir and interferon has no obvious therapeutic effect and some have serious side effects

Method used

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  • Phage display antibody library and monoclonal antibody against new coronavirus sars-cov-2 obtained based on its panning
  • Phage display antibody library and monoclonal antibody against new coronavirus sars-cov-2 obtained based on its panning
  • Phage display antibody library and monoclonal antibody against new coronavirus sars-cov-2 obtained based on its panning

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Embodiment 1

[0067] 1. Construction of phage-displayed antibody library

[0068] Take the synthetic antibody heavy chain variable region DNA (base sequence is SEQ ID NO: 1) as the template, use primers SfiVHback (base sequence is SEQ ID NO: 2) and VHFR2For (base sequence is SEQ ID NO: 3) ), amplify the HFR1-CDRH1-HFR2 gene fragment VH1 of the variable region VH of the antibody heavy chain by polymerase chain reaction, and use degenerate primer CDRH2libback (base sequence is SEQ ID NO: 4) and degenerate primer CDRH3lib9for (base The base sequence is SEQ ID NO: 5) Amplify the gene of CDRH2-HFR3-CDRH3-HFR4, namely VH2, perform agarose electrophoresis, confirm the amplified VH1 and VH2, cut out and purify the two gene fragments, and then use two gene sequences As a template, use primers SfiVHback (base sequence is SEQ ID NO: 2) and XhoVHfor (base sequence is SEQ ID NO: 6) to perform overlapping PCR;

[0069] The above PCR reactions used KOD-plus-neo (TOYOBO Bio), the reaction system was 50 μL...

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Abstract

The invention discloses a phage display antibody library and screened five antibodies capable of binding to the S protein of the new coronavirus SARS-CoV-2. The invention is based on synthetic biology and phage display technology, and introduces mutations into antibody variable regions. hypervariable region, and this gene was transferred to Escherichia coli, thereby constructing a 8 A synthetic antibody library of various antibodies; the phage display antibody library of the present invention can be screened to obtain antibodies with specificity and detection functions, which expands the powerful resources for biological research and medical diagnosis; the present invention has also screened five S strains that can follow the new coronavirus Protein-bound antibodies can be used for the detection of viruses, some antibodies can block the combination of viruses and cells, and have the ability to neutralize the infectivity of the new coronavirus, and can be used to prepare new coronavirus detection products, preparations for the preparation of new coronavirus drugs, and preparations for prevention or treatment Pharmaceutical preparations for diseases caused by the new coronavirus have broad application prospects.

Description

technical field [0001] The invention relates to the technical fields of biomedicine and molecular biology, in particular to a phage display antibody library and a monoclonal antibody against the new coronavirus SARS-CoV-2 obtained by panning based on the phage display antibody library. Background technique [0002] Compared with the original coronavirus, SARS-CoV-2 is not only contagious in severe patients, but also highly contagious in patients with mild symptoms and even in the incubation period, and there are many ways of transmission, which can be transmitted through droplets and contact. Great difficulty for prevention. [0003] Countries have proposed a variety of treatment options for the treatment of new coronary pneumonia, mainly using the small molecule drug Remdesivir, the combination of chloroquine and hydroxychloroquine, the combination of lopinavir and ritonavir, and the use of lopinavir and ritonavir. Unfortunately, the three-drug combination of Navir and int...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C40B40/02C12N15/10C12N15/70C12N15/13C07K16/10G01N33/577G01N33/569A61K39/42A61P31/14C12R1/92
CPCC07K16/10G01N33/577G01N33/56983A61P31/14C07K2317/56C07K2317/565C07K2317/76G01N2333/165G01N2469/10C07K2317/92C07K16/005Y02A50/30
Inventor 董金华董航单喜军
Owner 山东宽和正生物医药有限公司
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