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Adenovirus vector vaccine for preventing SARS-CoV-2 original strain and Beta strain

A carrier and vaccine technology, applied in the field of virus immunology, can solve the problems of decreased protective effect of Beta mutant strains and inability to achieve the immune effect of Beta strains, etc.

Active Publication Date: 2022-07-08
GUANGZHOU N BIOMED LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently marketed vaccines are inactivated vaccines, subunit protein vaccines, mRNA vaccines, and adenovirus vector vaccines. These vaccines are mainly aimed at the original strain of SARS-CoV-2, and their protective effects against Beta mutant strains are all reduced, so they cannot be used After immunization, it produces a very ideal immune effect on the Beta strain

Method used

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  • Adenovirus vector vaccine for preventing SARS-CoV-2 original strain and Beta strain
  • Adenovirus vector vaccine for preventing SARS-CoV-2 original strain and Beta strain
  • Adenovirus vector vaccine for preventing SARS-CoV-2 original strain and Beta strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Construction of the double antigen vector pAd5-S33 carrying the S gene of the Beta mutant strain and the RBD gene of the original strain

[0061] 1. Construction of the shuttle plasmid pGA1-S14c of the S gene of the Beta mutant strain

[0062] Using PGA1-NB14b (British mutant strain S gene vector, its sequence is obtained by point mutation on the original strain sequence of SEQ ID NO. Point mutation, which is obtained by adding the amino acids KV at positions 986 to 987 in the S2 region to PP; the above mutations promote the protein to maintain the pre-fusion conformation, thereby inducing neutralizing antibodies; the sequence of the NB14b is shown in SEQ ID NO: 2 ; preserved by Guangzhou Enbao Biomedical Technology Co., Ltd.) plasmid as template. The target fragment S14C-L was obtained by PCR amplification with Primer Star Mix (TaKaRa) using S-F and K417N-R as primers.

[0063] S14C-L amplification primer sequence:

[0064] S-F: ggtaccgagctcggatccgccaccatg...

Embodiment 2

[0102] Example 2 Adenovirus vector vaccine against SARS-CoV-2 original strain and Beta strain

[0103] 1. Rescue and production of Ad5-S33 vector

[0104] According to the conventional method, Ad5-S33 was linearized with PacI, recovered by ethanol precipitation, and transfected into 293 cells by cationic lipofection method. 4 hours after transfection, 2 ml of DMEM medium containing 5% fetal bovine serum was added, and incubated for 7 hours. -10 days, observe the cytopathic changes; after the poisoning, collect the cells and the culture supernatant, freeze and thaw 3 times in a 37-degree water bath and liquid nitrogen, and centrifuge to remove cell debris, and the supernatant infects a 10 cm dish; 2-3 days later, Collect cells and culture supernatant, freeze and thaw 3 times and centrifuge to remove cell debris, and infect 3-5 15 cm dishes with supernatant; after 2-3 days, collect cells, freeze and thaw 3 times and centrifuge to remove cell debris; supernatant After infecting ...

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Abstract

The invention belongs to the technical field of viral immunology, and discloses an adenovirus vector vaccine for preventing an SARS-CoV-2 original strain and a Beta strain, the S gene of the Beta strain and the RBD gene of the original strain are connected through efficient self-cleavage P2A, the antigen protein of the S protein of the Beta strain can be expressed, and the antigen of the RBD protein of the original strain can also be expressed. After immunizing an organism, the recombinant strain can generate a neutralizing antibody aiming at the SARS-CoV-2 of the Beta mutant strain, and also can generate a neutralizing antibody aiming at the original strain SARS-CoV-2, so that the organism is effectively protected from being infected by the Beta strain SARS-CoV-2 and the original strain SARS-CoV-2.

Description

technical field [0001] The invention belongs to the technical field of virus immunology, and in particular relates to an adenovirus vector vaccine for preventing the original strain and Beta strain of SARS-CoV-2. Background technique [0002] For the new coronavirus, taking precautions and blocking the spread of the virus is the key to controlling the epidemic. Vaccines are the most cost-effective intervention to prevent and control novel coronavirus infection. The published results of more than one hundred SARS-CoV-2 virus genome comparisons show that the overall degree of mutation of the virus is low and no recombination has occurred. Therefore, if the SARS-CoV-2 vaccine is successfully developed, it will be able to protect the population from the infection of the new coronavirus, thereby suppressing the outbreak of new epidemics. [0003] In the viral particle structure of the SARS-CoV-2 coronavirus, the S-protein (Spike protein, spike protein) that constitutes the "cro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/85C12N5/10A61K39/215A61P31/14C07K16/10G01N33/569
CPCC07K14/165C12N15/85A61K39/12A61P31/14C07K16/10G01N33/56983C07K2319/00C12N2770/20022C12N2770/20034G01N2333/165
Inventor 陈凌杨臣臣关素华刘波
Owner GUANGZHOU N BIOMED LTD
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