Quadrivalent SARS-CoV-2 chimeric nanoparticle vaccine as well as preparation method and application thereof

A nanoparticle and vaccine technology, applied in the field of biology, can solve the problems of serum immune escape, affect antigenicity, and low yield, and achieve the effects of improving immunogenicity, enhancing humoral immune response, and increasing expression yield

Pending Publication Date: 2022-06-24
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, most antibodies that neutralize viral infection are directed against the spike protein, making it an ideal antigen to elicit effective protective immunity against SARS-CoV-2 infection, but SARS-CoV- Mutations in 2 variants broadly affect their antigenicity, leading to widespread antibody and vaccine-induced seroimmune escape
The first generation of marketed vaccines, including mRNA, adenovirus vector vaccines, etc., are mainly designed and produced based on S-2P, but the low yield of S-2P expression, sensitivity to temperature and other factors limit the design of subunit vaccines based on S-2P Industrial production of vaccines

Method used

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  • Quadrivalent SARS-CoV-2 chimeric nanoparticle vaccine as well as preparation method and application thereof
  • Quadrivalent SARS-CoV-2 chimeric nanoparticle vaccine as well as preparation method and application thereof
  • Quadrivalent SARS-CoV-2 chimeric nanoparticle vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0066] The preparation method of the tetravalent chimeric nanoparticles of the present application comprises the following steps:

[0067] 1. Through computer-aided design such as Sic_axle and Rosetta, the fusion distance between the antigen and the nanoparticle carrier was determined, and the natural trimer domain of T4 phage fibrin was introduced at the same time, so as to determine the hinge for nanoparticle design in the sequence (linker) Length and antigen trimerization and selection of nanoparticle carriers based on the length design.

[0068] 2. Through the first host cell, different eukaryotic expression vectors were transferred into the first cell by transient transfection technology, respectively, to obtain different nanoparticle subunits of HexaPro-I53-50A1 (the first nanoparticle subunit to The fourth nanoparticle subunit), while using the second host cell, transform another I53-50B.4PT1 expression plasmid, and express after IPTG induction to obtain another nanopar...

Embodiment 1

[0071] Example 1 Construction, Expression and Purification of Chimeric Quadrivalent SARS-CoV-2 HexaPro Nanoparticle Vaccine

[0072] 1.1 Design of SARS-CoV-2 bicomponent antigen

[0073] Through computer-aided design such as Sic_axle and Rosetta, the HexaPro antigen (SARS-CoV-2) of SARS-CoV-2 wild type and VOCs (British strain, South African strain and Brazilian strain, corresponding to Alpha, Beta and Gamma types, respectively) was determined. The amino acid sequences of the wild-type, Alpha-type, Beta-type and Gamma-type HexaPro antigens are SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 in sequence) and the nanoparticle carrier (I53- 50A1) fusion gap; between the HexaPro antigen and the trimerized I53-50A1 subunit (SEQ ID NO: 5), the native trimer domain (SEQ ID NO: 6) of T4 phage fibrin was introduced, a segment comprising 16 A flexible linker of glycine-serine residues (SEQ ID NO: 7) and a rigid linker (SEQ ID NO: 8); for subsequent purification of the trimeri...

Embodiment 2

[0085] Example 2 Biophysical properties of chimeric tetravalent nanoparticle immunogens

[0086] 2.1 Particle size and distribution of nanoparticle immunogens

[0087] The four SARS-CoV-2 HexaPro-I53-50 NPs, Mosaic NPs and empty particles (I53-50 NPs) obtained in Example 1 were diluted with PBS solution to a concentration of 0.5 mg / mL; 50 μL were diluted The sample was added to a disposable solvent-resistant micro-test tube, and allowed to stand at 25°C for 2 min. The particle size of the nanoparticles was detected using a Zetasizer Ultra dynamic light scattering instrument (Malvern Panalytical), and the measurement angle was set to 173°, which was determined by measuring the intensity of scattered light. The size distribution of the protein was measured 5 times for each sample, and the average value was obtained to obtain the particle size.

[0088] like image 3 As shown in a, each nanoparticle immunogen is unimodal, the distribution maps of the four SARS-CoV-2 HexaPro-I53...

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Abstract

The invention belongs to the technical field of biology, discloses a tetravalent SARS-CoV-2 chimeric nanoparticle vaccine as well as a preparation method and application thereof, and particularly discloses a self-assembled nanoparticle which comprises a, at least one first nanoparticle subunit and b, a second nanoparticle subunit, the first nanoparticle subunit comprises a HexaPro protein and a first carrier subunit; the second nanoparticle subunit comprises a second carrier subunit; the first carrier subunit is I53-50A1, the second carrier subunit is I53-50B. 4PT1, and the HexaPro protein is connected with the first carrier subunit through a hinge. According to the self-assembled nanoparticles, HexaPro protein of the SARS-CoV-2 virus is displayed on the surfaces of the nanoparticles for the first time, higher antibody titer can be induced, and the self-assembled nanoparticles can be used for preventing SARS-CoV-2 virus infection.

Description

technical field [0001] The invention belongs to the technical field of biology, and in particular relates to a tetravalent SARS-CoV-2 chimeric nanoparticle vaccine and a preparation method and application thereof. Background technique [0002] At present, there are five Variants of Concerns (VOCs) in the world, namely B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), B.1.617.2 (Delta) and B.1.529 (Omicron), which have attracted widespread attention. Because they increase viral transmissibility and reduce neutralizing activity against vaccine-induced serum and monoclonal antibodies, they even cause immune escape, reinfection, and breakthrough infection. In addition, the rapidly increasing reports of emerging SARS-CoV-2 variants have created additional uncertainty about the effectiveness of existing vaccines, necessitating the development of new candidate multivalent vaccines targeting VOCs and transmitting variants. [0003] The SARS-CoV-2 spike protein plays a key role in vira...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K39/385A61K39/215A61P31/14
CPCC07K14/005A61K39/385A61K39/12A61P31/14C12N2770/20022C12N2770/20034C07K2319/735C07K2319/31C07K2319/02A61K2039/6031A61K2039/627
Inventor 曾木圣康银峰孙聪
Owner SUN YAT SEN UNIV CANCER CENT
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