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SARS-CoV-2 vaccine and preparation method thereof

A coronavirus, sars-cov-2 technology, applied in the field of vaccines, to achieve the effect of improving immunogenicity

Active Publication Date: 2020-06-02
CANSINO BIOLOGICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the S protein is the preferred antigen for the development of coronavirus vaccines, studies on SARS-CoV vaccines have shown that antibodies induced by the S protein can trigger antibody-dependent enhancement (antibody dependent enhancement, ADE); current research on the development of new coronavirus vaccines There are not many reports, but considering the high homology between the new coronavirus and SARS, whether the vaccine designed against the new coronavirus S protein can cause ADE is an important issue to be considered at the beginning of vaccine design

Method used

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

Examples

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Embodiment 1

[0072] The construction of the RBD-Linker-CTB fusion gene of embodiment 1 novel coronavirus (SARS-CoV-2)

[0073] The structure of the new coronavirus (SARS-CoV-2) and the composition of the coat protein are as follows: figure 1 shown. The S protein is the most important surface protein of the coronavirus. The S protein contains two subunits: S1 and S2. The receptor binding domain (RBD) is distributed in the S1 area and is responsible for recognizing cell receptors; the S protein can pass through the receptor binding area. Genetic recombination or mutation of (RBD) achieves transmission among different hosts and leads to higher lethality. More critically, coronaviruses recognize host cell receptors through RBD. Studies have shown that RBD is the neutralizing antibody epitope of S protein.

[0074] 1-1 Preparation of RBD-Linker-CTB fusion gene. The RBD-Linker-CTB fusion gene includes codon-optimized novel coronavirus (SARS-CoV-2) RBD gene, Linker and CTB gene. Among them, ...

Embodiment 2

[0085] The construction of the RBD-Linker-CRM197 (A) fusion gene of embodiment 2 novel coronavirus SARS-CoV-2

[0086] The specific construction method refers to Example 1. The RBD-Linker-CRM197(A) fusion gene includes the codon-optimized novel coronavirus (SARS-CoV-2) RBD gene, Linker and CRM197(A) genes.

[0087] Among them, the original sequence of the novel coronavirus (SARS-CoV-2) RBD gene is from NCBI and has been codon-optimized to increase the expression of the target antigen. The specific sequence is shown in SEQ ID NO:1. Wherein, the gene sequence of the Linker is GGGGS, as shown in SEQ ID NO:2. Wherein, CRM197(A) is the A region of CRM197, and the specific sequence is shown in SEQ ID NO:9.

[0088] In the following examples, the fusion genes prepared in Example 1 and Example 2 are collectively referred to as the RBD fusion gene of the novel coronavirus (SARS-CoV-2), referred to as the RBD fusion gene for short.

Embodiment 3

[0089] The development of embodiment 3 RBD-Linker-CTB fusion protein subunit vaccine

[0090] Transform Escherichia coli BL21 with the RBD-Linker-CTB fusion gene-pET9a expression vector, pick a single clone, recover in AMP-containing LB liquid medium, inoculate 100ml AMP-containing LB liquid medium for amplification, wait until OD 450 =0.5 (or in accordance with 0.4-0.6) Add 0.1 mM IPTG to induce expression. Induced expression results such as image 3 shown.

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Abstract

The invention relates to a preparation method for a vaccine capable of treating and / or preventing SARS-CoV-2 infection or COVID-19 diseases. The core antigen of the vaccine comprises the RBD (receptor binding zone) fusion protein of the SARS-CoV-2, and a vaccine form comprises an RBD fusion protein subunit vaccine, an RBD fusion protein mRNA vaccine or an RBD fusion protein adenovirus vector vaccine. The above vaccine immunizes an organism, and immune reaction for treating and / or preventing the SARS-CoV-2 infection can be generated so as to be used for treating and / or preventing COVID-19. The invention also relates to an RBD fusion gene, the RBD fusion protein, a carrier, a cell, a preparation method, a treatment method or a pharmacy purpose of the SARS-CoV-2.

Description

technical field [0001] The invention belongs to the technical field of vaccines, and in particular relates to designing the RBD fusion gene of a novel coronavirus (SARS-CoV-2) into an RBD fusion protein subunit vaccine, an RBD fusion protein mRNA vaccine, or an RBD fusion protein adenovirus vector vaccine. The vaccine can produce an immune response to prevent the new coronavirus (SARS-CoV-2) after immunizing the body, and is used to prevent the new coronavirus disease (COVID-19). Background technique [0002] Coronaviruses are non-segmented single-stranded positive-sense RNA viruses that belong to the Nidovirales Coronaviridae subfamily Orthocoronavirinae. According to serotype and genome characteristics, the Coronaviridae subfamily is divided into α , β, γ and δ four genera. So far, a total of 7 coronaviruses can infect humans: including 229E and NL63 of the genus α, OC43 and HKU1 of the genus β, Middle East respiratory syndrome-associated coronavirus (MERSr-CoV), severe a...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62A61K39/385A61K39/39A61K39/215A61P31/14
CPCA61K39/12A61K39/385A61K39/39A61K2039/5256A61K2039/53A61K2039/55544A61K2039/55561A61K2039/6037A61P31/14C07K14/005C07K2319/02C07K2319/40C07K2319/55C12N2770/20022C12N2770/20034
Inventor 李军强司伟雪崔海燕李荩巢守柏朱涛
Owner CANSINO BIOLOGICS INC
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