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A vaccine against sars-cov-2

A technology of polynucleotides and expression vectors, applied in the field of vaccines against SARS-CoV-2, can solve the problems of lack of SARS-CoV-2 antigen preparations and achieve good immunogenicity

Active Publication Date: 2022-01-25
重庆博唯佰泰生物制药有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In view of the above-mentioned shortcoming of prior art, the object of the present invention is to provide a kind of vaccine against SARS-CoV-2, for solving the problem of lacking effective SARS-CoV-2 antigen preparation in the prior art

Method used

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  • A vaccine against sars-cov-2
  • A vaccine against sars-cov-2
  • A vaccine against sars-cov-2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1 Construction of S protein engineering strains with different lengths of N-terminal and C-terminal truncation

[0102] 1. Selection of expressed genes and optimal design of codons

[0103] The amino acid sequence encoding S protein with different lengths truncated at the N-terminus and C-terminus refers to the Chinese strain 2019-nCoV_HKU-SZ-002a_2020 (GenBank accession number MN938384.1). The protein sequence of RBD193 is shown in SEQ ID NO: 1; the protein sequence of RBD194 As shown in SEQ ID NO: 2; RBD219 protein sequence is shown in SEQ ID NO: 3; RBD220 protein sequence is shown in SEQ ID NO: 4; RBD237 protein sequence is shown in SEQ ID NO: 5; RBD258 protein sequence is shown in SEQ ID NO: 6; RBD263 protein sequence is shown in SEQ ID NO: 7; RBD290 protein sequence is shown in SEQ ID NO: 8; RBD301 protein sequence is shown in SEQ ID NO: 9; RBD333 protein sequence is shown in SEQ ID NO 10; RBD343 protein sequence as shown in SEQ ID NO: 11; RBD386 protein s...

Embodiment 2

[0173] The preparation technology of embodiment 2 RBD194, RBD219, RBD220

[0174] 1. Fermenter culture of RBD194, RBD219, RBD220

[0175] The RBD194, RBD219, and RBD220 expression strains stored in glycerol tubes were inoculated in YPG medium at a ratio of 1:1000 (v / v), and cultured in shake flasks for 16-24 hours. Inoculate it into the fermentation medium (Basic salts) at a ratio of 1:20 (v / v), maintain the dissolved oxygen value by adjusting the rotation speed, tank pressure and ventilation, and add glycerol to grow the yeast cells. After the consumption of glycerol is over, methanol is added for induction, and the dissolved oxygen value is maintained by adjusting the rotation speed, tank pressure and ventilation. After 60-70 hours of induction, the fermentation is terminated. After centrifugation, the fermentation supernatant was collected for subsequent purification. The fermentation supernatants at different times were taken for SDS-PAGE identification (Fig. 12-14). The...

Embodiment 3

[0178] Example 3 Detection of receptor binding properties of RBD194, RBD219, and RBD220

[0179] 1. In vitro receptor binding verification

[0180] A 96-well plate was coated with 0.2 μg soluble ACE2 (Nanjing GenScript Biotech), placed overnight at 4°C, and washed 5 times with PBST. Add blocking solution (PBST containing 5% milk powder) to each well and place at 37°C for 2 hours. Wash the plate 5 times with PBST. Add 0.05 μg and 0.005 μg of RBD194, RBD219, and RBD220 samples to the microtiter plate, and incubate at room temperature for 1 hour. Wash the plate 5 times with PBST. Add 100 μl 1:2500 dilution of SARS-CoV-2S1 rabbit monoclonal antibody (Sino Biological) to each well and incubate at room temperature for 1 hour. Wash the plate 5 times with PBST. Add 100 μl 1:1000 diluted goat anti-rabbit IgG-HRP (Beijing Dingguo) to each well and incubate at room temperature for 1 hour. Wash the plate 5 times with PBST. Add 100 μl of freshly prepared color development solution (...

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Abstract

The present invention provides a vaccine against SARS‑CoV‑2. The present invention adopts the S protein of SARS-CoV-2 after transformation, especially the amino acid sequence such as SEQ ID NO: 2, such as the protein shown in SEQ ID NO: 3 or SEQ ID NO: 4, is used to prepare vaccine preparations, and has better Its immunogenicity provides a new solution for the treatment of diseases caused by SARS‑CoV‑2.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a vaccine against SARS-CoV-2. Background technique [0002] Coronavirus (coronavirus) belongs to the Coronaviridae (cirinaviridae) of Nidovirales. According to the genome homology, it can be divided into 4 genera: α, β, γ, δ, and the β genus can also be Divided into 4 departments - a, b, c, d. Coronaviruses can infect a variety of vertebrates, including humans, mice, pigs, bats, cats, cows, and chickens. A variety of coronaviruses that infect humans have been discovered for decades, including HCoV-OC43 and HCoV-229E isolated from the UK in the 1960s, SARS-CoV that broke out in China in 2002, and HCoV-NL63 that appeared in the Netherlands in 2004 , HCoV-HKU1 isolated from Hong Kong, China in 2005, and MERS-CoV that broke out in the Middle East in 2012. Most of these viruses cause respiratory diseases. Among them, SARS-CoV and MERS-CoV caused the severe acute respiratory syndrome in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/165C12N15/50C12N15/81C12N1/19A61K39/215A61P31/14C12R1/865C12R1/78C12R1/84C12R1/645
CPCC07K14/005C12N15/81C12N15/815A61K39/215A61P31/14C12N2770/20022C12N2770/20034
Inventor 傅文彬丁珊高兴张千里沈琼
Owner 重庆博唯佰泰生物制药有限公司
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