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SARS-CoV-2 RBD Conjugated Nanoparticle Vaccine

A nanoparticle and vaccine technology, applied in the field of immunomedicine, can solve problems such as existing needs, and achieve the effects of easy production, high yield, and safe use of vaccines

Active Publication Date: 2021-03-02
SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, a variety of SARS-CoV-2 vaccines have been developed around the world, mainly including inactivated vaccines, subunit vaccines, viral vector vaccines and nucleic acid vaccines, and 7 vaccines have been approved to enter the third phase of clinical trials, but they have not yet been marketed to prevent A vaccine against SARS-CoV-2, so there is still a need for developing a preventive vaccine that produces high titers of neutralizing antibodies against SARS-CoV-2

Method used

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  • SARS-CoV-2 RBD Conjugated Nanoparticle Vaccine
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  • SARS-CoV-2 RBD Conjugated Nanoparticle Vaccine

Examples

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preparation example Construction

[0090] The present invention also relates to a method for preparing the immunogenic complex as described above, comprising:

[0091] The immunogenic complex is obtained by expressing the fusion proteins of a) component and b) component, co-incubating after purification, and self-assembling.

[0092] According to yet another aspect of the present invention, it also relates to the application of the above-mentioned immunogenic complex, or the above-mentioned nanoparticle vaccine in the preparation of drugs for the treatment of novel coronavirus pneumonia.

[0093] The present invention further provides a method for protecting a subject from SARS-CoV-2 virus infection, which comprises administering to said animal an effective amount of the nanoparticle vaccine according to the present invention.

[0094] The subjects for the above purposes can refer to patients or animals suspected of carrying SARS-CoV-2, especially mammals, such as bats and civet cats; preferably primates, more ...

Embodiment 1

[0102] Embodiment 1. Design based on SARS-CoV-2 RBD nanoparticle vaccine

[0103] This example describes the design of Ferritin (24-mer), mi3 (60-mer) and I53-50 (120-mer) nanoparticles based on the SARS-CoV-2 RBD protein.

[0104] Considering that different nanoparticles can display different copy numbers of antigens on their surface, this example designs three SARS-CoV-2 RBD-conjugated nanoparticles: RBD-Ferritin, RBD-mi3 and RBD-I53-50.

[0105] Ferritin (SEQ ID NO: 2) is an octahedron composed of 24 subunits. Ferritin protein is the N-terminal 2-9 residues of the lower subunit of bullfrog (Rana catesbeiana) ferritin (UniProt: P07797) fused to 3-167 residues of Helicobacter pylori non-heme ferritin by molecular biological means. In order to eliminate the influence of the N-terminal potential glycosylation site, the present invention adopts N8Q and N19Q point mutations at residues 8 and 19 of bullfrog ferritin. To preserve the salt bridge between residues 6R and 14E of H. ...

Embodiment 2

[0109] Example 2. Expression and purification based on SARS-CoV-2 RBD nanoparticle vaccine

[0110] 1. Experimental materials

[0111] (1) Vectors and strains required for constructing recombinant vectors: mammalian expression vector VRC8400, Escherichia coli expression vector modified PET-28a+, Escherichia coli competent DH5a cells, Rosseta cells.

[0112] Protein expression cell line: HEK293-F cells (derived from human embryonic kidney epithelial cells).

[0113] (2) Reagents and consumables: PCR enzyme and recombinase (purchased from Novizyme Co., Ltd.), endonuclease (purchased from NEB), cell transfection reagent PEI-MAX (Polysciences, Inc., Cat. No. 24765- 1) Mammalian cell culture medium Union293 medium (purchased from Shanghai Yonglian Biology), histidine-tagged protein purification agarose magnetic beads (purchased from GE), and other conventional reagents and consumables are commercially available.

[0114] (3) Genes: △N1-SpyCatcher-Ferritin (SEQ ID NO: 18), △N1-Spy...

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Abstract

The invention relates to the field of immunomedicine, in particular to a SARS‑CoV‑2 RBD conjugated nanoparticle vaccine. The vaccine comprises an immunogenic complex comprising: a) a nanoparticle carrier obtained by self-assembly of a carrier protein fused with SpyCatcher; b) RBD of a SARS‑CoV‑2 virus fused with SpyTag Antigen; the carrier protein is selected from Ferritin, mi3 and I53-50; the carrier protein and the antigen are covalently linked by SpyCatcher-SpyTag.

Description

technical field [0001] The invention relates to the field of immunomedicine, in particular to a SARS-CoV-2 RBD conjugated nanoparticle vaccine. Background technique [0002] Severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) is a pathogen that causes acute lower respiratory tract infection with a high fatality rate, and can cause diseases such as novel coronavirus pneumonia (COVID-19). Due to the strong infectivity, fast transmission speed and high lethality of SARS-CoV-2, there is no effective specific drug and vaccine to prevent and control it. Therefore, the development of a preventive vaccine against this virus is particularly urgent. [0003] The S protein is a trimeric glycoprotein, a fusion protein of class I viruses, which also includes HIV glycoprotein 160 (Env), influenza hemagglutinin (HA), paramyxovirus F and Ebola virus glycoproteins. The S protein can bind to the virus receptor of the host cell, and is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/215A61K39/385A61P31/14C07K19/00
CPCA61K39/12A61K39/385A61P31/14C07K14/005C07K14/47C12N2770/20034A61K2039/6031A61K2039/627C12N2770/20022C07K2319/735
Inventor 曾木圣康银峰孙聪冯国开
Owner SUN YAT SEN UNIV
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