Novel recombinant coronavirus protein vaccine as well as preparation method and application thereof

A new type of coronavirus technology, which is applied in the field of recombinant new coronavirus protein vaccines, can solve problems such as concerns about the effectiveness of new crown vaccines, and achieve the effects of vaccine production capacity advantages and economic cost advantages

Pending Publication Date: 2022-04-12
NAT VACCINE & SERUM INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The emergence of these strains with strong immune escape ability has caused great concern about the effectiveness of the existing new crown vaccine

Method used

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  • Novel recombinant coronavirus protein vaccine as well as preparation method and application thereof
  • Novel recombinant coronavirus protein vaccine as well as preparation method and application thereof
  • Novel recombinant coronavirus protein vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1: Recombinant novel coronavirus RBD trimeric protein based on protein structure and computational biology design

[0100]The new coronavirus continues to mutate, and the emergence of multiple mutant strains has led to multiple rounds of outbreaks. According to the different transmissibility, pathogenicity or immune escape ability of the mutant strains, the World Health Organization classifies five mutant strains as variants of concern (VOCs), including Alpha (B.1.1.7) , Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), and Omicron (B.1.1.529). Among the five VOCs, a large number of studies have confirmed that the Beta variant has a strong immune escape ability, and the Delta variant has a strong transmission ability. Preliminary evidence shows that the Omicron mutant strain has both strong transmission ability and strong immune evasion ability, and its immune evasion ability is even much higher than that of the Beta strain. The emergence of these strains with s...

Embodiment 2

[0105] Example 2: Recombinant protein expression, purification and identification

[0106] According to the codon bias of the CHO cell expression system, the nucleotide sequence encoding the recombinant protein C05G11 (amino acid sequence shown in SEQ ID NO.1) was codon optimized, and the optimized nucleotide sequence was shown in SEQ ID NO. 2. The CHO cell expression vector was constructed and then transfected into 293FT cells or CHO cells to construct a recombinant cell line. The cell line capable of stably secreting and expressing the recombinant protein C05G11 was screened by the limiting dilution method. After cell culture, the supernatant was harvested and purified by serial chromatography Finally, the recombinant protein C05G11 with a purity ≥ 95% was obtained. SDS-PAGE detection results such as figure 2 As shown, the molecular weight of the protein is 70-100kD, and some product-related substances can be seen at the same time, such as dimer protein and monomer protei...

Embodiment 3

[0108] Example 3: Detection of Physicochemical Properties and Biological Activity of C05G11 Protein

[0109] Purified C05G11 protein, C05 protein (amino acid sequence shown in SEQ ID No.23, obtained through recombinant expression and chromatography purification of 293FT cells or CHO cells) and C05C protein (amino acid sequence shown in SEQ ID No.24, Recombinantly expressed in 293FT cells or CHO cells and purified by chromatography), the RBD protein of the prototype strain (manufacturer: Beijing Yiqiao Shenzhou Technology Co., Ltd.; article number: 40592-V08B), the RBD protein with the same mutation site as the Beta strain virus (K417N , E484K, N501Y; manufacturer: Beijing Yiqiao Shenzhou Technology Co., Ltd.; article number: 40592-V08H85), RBD protein (L452R, T478K; manufacturer: Beijing Yiqiao Shenzhou Technology Co., Ltd.; article number: 40592-V02H3), the RBD protein (G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H) co...

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PUM

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Abstract

The invention discloses a novel recombinant coronavirus protein, the novel recombinant coronavirus protein covers specific mutation sites of Delta, Beta and Omicron variant RBD, a recombinant vaccine using the novel recombinant coronavirus protein as a target antigen has a broad-spectrum protection capability of trans-epidemic strains, and in addition, compared with a traditional method for preparing a plurality of monovalent vaccines, the novel recombinant coronavirus protein has a broad-spectrum protection capability of trans-epidemic strains. Compared with the strategy of realizing broad-spectrum protection through multivalent combination, the multivalent broad-spectrum protection effect is realized on one antigen molecule, and the vaccine has obvious advantages in the aspects of time cost and economic cost of vaccine preparation and vaccine productivity.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a recombinant novel coronavirus protein vaccine, its preparation method and application. Background technique [0002] Novel coronavirus (SARS-CoV-2), belonging to Nidovirales, Coronaviridae, Orthocoronavirus subfamily, Betacoronavirus genus, Sarbecovirus subgenus, SARS-like virus species, single-stranded positive-sense RNA virus , with an envelope, the full length of the genome is about 29.9kb, most of which encode non-structural proteins, involved in virus replication and translation, and a small part of the sequence encodes structural proteins, such as: S protein (spike protein,), M protein (membrane protein), E protein (envelope protein) and N protein (nucleo protein). In addition, there are several accessory proteins: 3a, 3b, p6, 7a, 7b, 8b, 9b and orf14, which are all involved in virus assembly. The S, M and E proteins constitute the viral envelope and are the main surface anti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C07K19/00C12N15/50C12N15/62C12N15/85A61K39/215A61P31/14
Inventor 李启明梁宇苏计国侯俊伟韩子泊张靖张学峰靳玉琴刘兆明杜丽芳雷泽华唐芳张浩邵帅侯亚楠马智静陈实郑凡刘宁沈福杰
Owner NAT VACCINE & SERUM INST
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