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Phage display antibody library and monoclonal antibodies aiming at novel coronavirus SARS-CoV-2 and obtained by panning based on same

A phage display, monoclonal antibody technology, applied in the fields of biomedicine and molecular biology, can solve problems such as toxic side effects, prevention difficulties, no obvious treatment effect, etc., to achieve the effect of expanding powerful resources

Active Publication Date: 2020-10-16
山东宽和正生物医药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 antibodies aiming at novel coronavirus SARS-CoV-2 and obtained by panning based on same
  • Phage display antibody library and monoclonal antibodies aiming at novel coronavirus SARS-CoV-2 and obtained by panning based on same
  • Phage display antibody library and monoclonal antibodies aiming at novel coronavirus SARS-CoV-2 and obtained by panning based on same

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Embodiment 1

[0067] 1. Construction of phage display antibody library

[0068] Using the synthetic antibody heavy chain variable region DNA (base sequence as SEQ ID NO: 1) as a template, use primers SfiVHback (base sequence as SEQ ID NO: 2) and VHFR2For (base sequence as SEQ ID NO: 3 ), amplify the HFR1-CDRH1-HFR2 gene fragment VH1 of the antibody heavy chain variable region VH 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, that is, VH2, perform agarose electrophoresis, confirm the amplified VH1 and VH2, cut out and purify the two gene fragments, and then use the two gene sequences As a template, perform overlapping PCR with primers SfiVHback (base sequence is SEQ ID NO: 2) and XhoVHfor (base sequence is SEQ ID NO: 6);

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

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Abstract

The invention discloses a phage display antibody library and five strains of screened antibodies capable of being combined with S protein of novel coronavirus SARS-CoV-2. Mutation is introduced into an ultra-variable region of an antibody variable region based on synthetic biology and a phage display technology, and a gene is transferred into escherichia coli, so that a synthetic antibody librarycontaining 108 kinds of antibodies is constructed; the phage display antibody library provided by the invention can perform screening to obtain the antibody with the specificity and the detection function, so that powerful resources of biological research and medical diagnosis are expanded; and the five strains of antibodies capable of being combined with the S protein of the novel coronavirus arefurther screened out and can be used for detecting the virus, part of the antibodies can block combination of the virus and cells, and the antibodies have the capacity of neutralizing novel coronavirus infectivity, can be used for preparing a novel coronavirus detection product, preparing a drug for inhibiting the novel coronavirus and preparing a pharmaceutical preparation for preventing or treating diseases caused by the novel coronavirus, and have a wide application prospect.

Description

technical field [0001] The present invention relates to the technical fields of biomedicine and molecular biology, specifically a phage display antibody library and a monoclonal antibody against the new coronavirus SARS-CoV-2 obtained by panning the phage display antibody library. Background technique [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 has brought great difficulties to prevention. [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 o...

Claims

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Application Information

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Patent Type & Authority Applications(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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