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Three recombinant adenoviruses, RBD of SARS-CoV-2 Spike protein and application of three recombinant adenoviruses and RBD of SARS-CoV-2 Spike protein

A recombinant adenovirus, adenovirus technology, used in recombinant DNA technology, polypeptides containing Strep tags, applications, etc.

Pending Publication Date: 2021-11-02
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are no specific drugs and vaccines for the treatment and prevention of this disease

Method used

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  • Three recombinant adenoviruses, RBD of SARS-CoV-2 Spike protein and application of three recombinant adenoviruses and RBD of SARS-CoV-2 Spike protein
  • Three recombinant adenoviruses, RBD of SARS-CoV-2 Spike protein and application of three recombinant adenoviruses and RBD of SARS-CoV-2 Spike protein
  • Three recombinant adenoviruses, RBD of SARS-CoV-2 Spike protein and application of three recombinant adenoviruses and RBD of SARS-CoV-2 Spike protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1, preparation and identification of recombinant virus

[0079] 1. Construction of recombinant plasmids

[0080] pAdC68XY4 pAdC68-ΔE1(GFP)-ΔE3(SwaI)-E4(orf3-6Hu5) vector is a circular plasmid, as shown in sequence 7 of the sequence listing. pAdC68XY4 The pAdC68-ΔE1(GFP)-ΔE3(SwaI)-E4(orf3-6Hu5) vector is derived from Simian adenovirus 25 (NCBI Reference Sequence: AC_000011.1), and has been modified as follows: Parts of E1 and E3 have been deleted, E1 Deletion of part of the region prevents the generated recombinant virus from replicating in ordinary cells to enhance its biological safety, and part of the deletion of the E3 region increases the insertion capacity of foreign genes. The pAdC68XY4 pAdC68-ΔE1(GFP)-ΔE3(SwaI)-E4(orf3-6Hu5) vector has two SrfI restriction recognition sequences, and the small fragment between the two restriction restriction recognition sequences has the EGFP gene. When using the pAdC68XY4 pAdC68-ΔE1(GFP)-ΔE3(SwaI)-E4(orf3-6Hu5) vecto...

Embodiment 2

[0124] Embodiment 2, the application of recombinant virus

[0125] The virus liquids used for animal immunization were all the virus liquids prepared in Example 1.

[0126] 1. Animal immunity

[0127] 6-week-old female BALB / C mice were divided into 24 groups, with 5 mice in each group, and were treated as follows:

[0128] The first group (G1 group): a single immunization was carried out by nasal drops, and the immune substance was ChAdTS virus liquid (the amount of virus given to a single mouse was 2×10 8 vp);

[0129] The second group (G2 group): a single immunization was carried out by nasal drop method, and the immune substance was ChAdTS virus liquid (the amount of virus given to a single mouse was 2×10 9 vp);

[0130] The third group (G3 group): a single immunization was carried out by nasal drop method, and the immunization material was ChAdTS virus liquid (the amount of virus given to a single mouse was 2×10 10 vp);

[0131] The fourth group (G4 group): a single ...

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Abstract

The invention discloses three recombinant adenoviruses, RBDs of SARS-CoV-2 Spike protein and application of the three recombinant adenoviruses and the RBDs of the SARS-CoV-2 Spike protein. The invention provides a recombinant adenovirus: a recombinant adenovirus expressing a protein represented by a sequence 3 in a sequence table; a recombinant adenovirus expressing the protein represented by the sequence 5 of the sequence table; and the recombinant adenovirus expresses the protein shown in the sequence 1 of the sequence table. The protein as shown in the sequence 1 in the sequence table is the full-length SARS-CoV-2 Spike protein. The protein as shown in the sequence 3 in the sequence table is the SARS-CoV-2 receptor binding domain RBD protein with the membrane anchoring signal peptide. The protein as shown in the sequence 5 in the sequence table is the SARS-CoV-2 receptor binding domain RBD protein with the secretory signal peptide. The vaccine aiming at the novel coronavirus SARS-CoV-2, which is researched and developed by the invention, has an important theoretical guidance value and a wide application prospect, and provides possibility for radical treatment of novel coronavirus pneumonia.

Description

technical field [0001] The present invention relates to three kinds of recombinant adenoviruses and the RBD of SARS-CoV-2 Spike protein and their application. Background technique [0002] Most patients with SARS-CoV-2 infection develop severe respiratory disease, and the clinical symptoms are very similar to the disease caused by SARS-CoV that broke out in 2003. Because the disease can be transmitted from person to person, it has attracted great attention around the world. So far, there are no specific drugs and vaccines for the treatment and prevention of this disease. [0003] The current outbreak of novel coronavirus pneumonia and the outbreak of SARS in 2002 were both caused by previously unknown coronaviruses (Coronavirus, CoV). Due to its unpredictability, coronavirus can be transmitted through droplet transmission, contact with respiratory secretions, etc., causing serious consequences, and has become one of the major threats to human health. Contents of the inve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/861C07K14/165C07K19/00C12N15/62C12N15/50A61K39/215A61P31/14A61P11/00
CPCC12N7/00C12N15/86C07K14/005A61K39/12A61P31/14A61P11/00C12N2710/10043C12N2770/20022C07K2319/02C07K2319/21C07K2319/22C07K2319/43C07K2319/41C12N2770/20034
Inventor 张林琦周东明李明茜史宣玲
Owner TSINGHUA UNIV
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