Adenovirus vector recombinant new coronavirus B.1. 1.529 variant vaccine and application thereof

An adenovirus and mutant technology, applied in the field of bioengineering, can solve the problems of reduction, the failure protection of the new crown prototype strain vaccine, etc., and achieve the effect of high-efficiency expression

Active Publication Date: 2022-04-26
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of mutations accumulated in the spike protein enable the epidemic strain to escape the antibodies elicited by the new crown prototype strain vaccine to a certain exte

Method used

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  • Adenovirus vector recombinant new coronavirus B.1. 1.529 variant vaccine and application thereof
  • Adenovirus vector recombinant new coronavirus B.1. 1.529 variant vaccine and application thereof
  • Adenovirus vector recombinant new coronavirus B.1. 1.529 variant vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Comparison before and after optimization of the nucleotide sequence of the new crown B.1.1.529 mutant strain antigen

[0035] We selected the mutant strain B.1.1.529 (GISAID Accession ID: EPI_ISL_6640917) as a template to obtain the S protein sequence of the mutant strain. Combined with empirical optimization, the nucleotide sequence shown in SEQ ID NO.1 was obtained.

[0036] The length of the original spike protein sequence is 3813 bp, of which 1129 bases are A, accounting for 29.61%, C is 716, accounting for 18.78%, G is 702, accounting for 18.41%, and T is 1266, accounting for 33.2% ( figure 1 ). After empirical optimization, the sequence length is 3837bp, A is 912, accounting for 23.77%, C is 1185, accounting for 30.88%, G is 1003, accounting for 26.14%, T is 737, accounting for 19.21% ( figure 2 ), the GC base content increased from 37.19% to 57.02%. Since AT pairing generates 2 hydrogen bonds and GC pairing generates 3 hydrogen bonds, the increase ...

Embodiment 2

[0039] Example 2: Recombinant virus vaccine packaging and in vitro expression identification

[0040] During the gene synthesis process, the synthesized product was cloned into the pDC316 vector (Microbix BiosystemsInc.), and the map of the obtained plasmid (pDC316-nCoV-B.1.1.529) was as follows Figure 7 shown. HEK293 cells were co-transfected with the pDC316-nCoV-B.1.1.529 plasmid and the adenovirus backbone plasmid pBHGlox_E1, 3Cre (Microbix Biosystems Inc.), and maintained in DMEM medium containing 5% FBS until cytopathic. During the maintenance culture process, the pDC316 vector contains the left inverted repeat sequence of adenovirus, packaging signal sequence and partial fragments of the antigen gene, relying on Cre / Loxp site-specific recombination and splicing of the virus backbone sequence to form a complete genome of the virus, start Progeny virus synthesis and assembly. With the continuous synthesis of the progeny virus, the cell disease gradually increased. After...

Embodiment 3

[0042] Example 3: Evaluation of Immune Response of New Crown B.1.1.529 Variant Vaccine

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Abstract

The invention provides a novel coronavirus B.1. 1.529 variant vaccine taking a human type 5 replication-deficient adenovirus as a vector. The invention further provides a preparation method of the novel coronavirus B.1. 1.529 variant vaccine. On the premise that an expression protein main body is still novel coronavirus B.1. 1.529 variant spike protein, a recombinant virus vector vaccine prepared from a nucleotide sequence after experience optimization can effectively stimulate an organism to generate a binding antibody, a neutralizing antibody and cellular immune response aiming at the B.1. 1.529 variant virus after immunization, and has good immunogenicity.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a recombinant novel coronavirus B.1.1.529 variant vaccine. Background technique [0002] The new coronavirus is an RNA virus, which is very prone to mutations during the continuous transmission process, and naturally screens out the dominant mutant strains with enhanced transmission capabilities. A mutant strain of Omicron (B.1.1.529) was discovered for the first time in November 2021. On November 26, the World Health Organization defined it as the fifth "mutant strain of concern". After December 2021, the mutant strain began to spread widely around the world, and by the end of February 2022, the coverage rate of the B.1.1.529 mutant strain was close to 100%. [0003] Compared with the prototype strain of the novel coronavirus, the B.1.1.529 mutant strain's Spike protein (Spike, S) contains more than 37 mutation sites, of which about 15 mutation sites are loca...

Claims

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Application Information

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IPC IPC(8): C12N15/50C12N15/861A61K39/215A61P31/14A61P11/00
CPCC07K14/005C12N15/86A61K39/12A61P31/14A61P11/00C12N2770/20022C12N2770/20034C12N2710/10343C12N2800/107C12N2800/22A61K2039/53A61K2039/5256A61K2039/543A61K2039/54A61K2039/575A61K2039/57
Inventor 陈薇王步森侯利华徐婧含吴诗坡张哲赵拯浩张金龙宋小红王玉东陈旖付玲
Owner ACADEMY OF MILITARY MEDICAL SCI
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