SARS-CoV-2 coronavirus vaccine and preparation method thereof

A coronavirus, sars-cov-2 technology, applied in the field of biological genetic engineering, to achieve the effect of high biological activity, long-lasting immunogenicity and long half-life

Active Publication Date: 2021-04-09
GUANGZHOU DOUBLLE BIOPRODUCT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there is no research report on the preparation of a vaccine against SARS-CoV-2 viral S protein truncated by adenoviral vector

Method used

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  • SARS-CoV-2 coronavirus vaccine and preparation method thereof
  • SARS-CoV-2 coronavirus vaccine and preparation method thereof
  • SARS-CoV-2 coronavirus vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The construction of embodiment 1SARS-CoV-2 coronavirus vaccine Ad / S1

[0069] (1) Amplification of the S gene fragment:

[0070] Using the S gene obtained in the basic example as a template, amplify with the following primers:

[0071] V1 (SEQ ID NO: 1): TCC CCCGGG ATGTTCGTCTTCCTGGTCCT

[0072] V4 (SEQ ID NO: 4): CCC AAGCTT TTACCGGGCTCTTCTGGGAGAGT

[0073] The S1 gene is amplified, that is, the positions 1-2055 of the S gene of the basic example, and the amplified product is named S1.

[0074] (2) Construction of recombinant plasmids:

[0075] The S1 fragment of the PCR amplification product obtained above was identified by gel running and recovered by gel cutting, and digested with SmaI and HindIII at 37°C. Digest pShuttle with these two enzymes at the same time. Next, use T4 ligase to ligate the digested PCR product with pShuttle overnight at 16°C. The ligation product was transformed into Escherichia coli DH5α, positive clones were screened by ampicillin res...

Embodiment 2

[0078] The construction of embodiment 2SARS-CoV-2 coronavirus vaccine Ad / S1N

[0079] The difference with Example 1 is: the PCR amplification primers used in step (1) are:

[0080] V1 (SEQ ID NO: 1): TCC CCCGGG ATGTTCGTCTTCCTGGTCCT

[0081] V3 (SEQ ID NO: 3): CCC AAGCTT TTAGGCCACCTGGTTGCTTGTAT

[0082] The rest is the same.

[0083] The amplified product is named S1N fragment, and the final SARS-CoV-2 vaccine is named Ad / S1N vaccine, and the inserted gene is sequenced, and the sequencing results are shown in Figure 4 .

Embodiment 3

[0084] The construction of embodiment 3SARS-CoV-2 coronavirus vaccine Ad / S1C

[0085] The difference with Example 1 is: the PCR amplification primers used in step (1) are:

[0086] V2 (SEQ ID NO: 2): TCC CCCGGG ATGAGGGTGCAGCCAACCGAG

[0087] V4 (SEQ ID NO: 4): CCC AAGCTT TTACCGGGCTCTTCTGGGAGAGT

[0088] The rest is the same.

[0089] The amplified product is named S1C fragment, and the final SARS-CoV-2 vaccine is named Ad / S1C vaccine, and the inserted gene is sequenced, and the sequencing results are shown in Figure 5 .

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Abstract

The invention discloses a SARS-CoV-2 coronavirus vaccine and a preparation method thereof. An S gene of a SARS-CoV-2 coronavirus is subjected to codon optimization, truncated body and mutant sequences of the S gene are introduced into a secretory defective adenovirus vector, and packaging is conducted to obtain a corresponding recombinant adenovirus; and the recombinant adenovirus can express SARS-CoV-2 virus related protein in vivo, processing, folding, glycosylation and other modifications are completed, the native conformation of S protein is basically maintained, and the characteristics of high biological activity, long half-life period, lasting immunogenicity and the like are achieved. According to the vaccine and the method, by adopting the defective adenovirus vector carrying secretory peptide, a recombinant adenovirus vaccine can be secreted out of cells after being expressed in vivo, so that the humoral immunity is activated.

Description

technical field [0001] The invention belongs to the field of biological genetic engineering, in particular to a SARS-CoV-2 coronavirus vaccine. Background technique [0002] The new coronavirus has a strong ability to spread from person to person, and the transmission index R0 is about 2.5. The virus mainly infects people through the respiratory tract or contact transmission. It can cause acute pneumonia and can also cause serious damage to other systems such as the urinary system, digestive system and nervous system. Currently, there is no effective drug treatment. Therefore, it is very important to design and develop an effective vaccine against this virus. [0003] In addition, the SARS-CoV-2 coronavirus is a single-stranded RNA virus, which is very unstable. It is subject to immune supervision and rejection in somatic cells, and it is very prone to mutations. Therefore, vaccines designed based on antigenic determinants with relatively conserved sequences can induce rel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61P31/14A61P11/00C12N15/50C12N15/861C07K14/165
CPCA61K39/12A61P31/14A61P11/00C07K14/005C12N15/86A61K2039/5256C12N2800/22C12N2770/20022C12N2770/20034C12N2710/10043Y02A50/30
Inventor 黄文林田烁周晓鸿
Owner GUANGZHOU DOUBLLE BIOPRODUCT CO LTD
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