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SARS-CoV-2 polypeptide vaccine using papillomavirus (PV) virus-like particle (VLP) presentation antigen

A virus-like particle, coronavirus technology, applied in the field of novel coronavirus polypeptide vaccines, which can solve the problems of no antiviral drugs or regulatory approval of vaccines

Active Publication Date: 2021-02-02
BEIJING HEALTH GUARD BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no antiviral drugs or vaccines for COVID-19 that have received regulatory approval

Method used

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  • SARS-CoV-2 polypeptide vaccine using papillomavirus (PV) virus-like particle (VLP) presentation antigen
  • SARS-CoV-2 polypeptide vaccine using papillomavirus (PV) virus-like particle (VLP) presentation antigen
  • SARS-CoV-2 polypeptide vaccine using papillomavirus (PV) virus-like particle (VLP) presentation antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Example 1: Design and synthesis of codon-optimized genes

[0117] Referring to the codon preference of Escherichia coli for gene transcription, the coding sequence of the SARS-CoV-2 spike protein epitope polypeptide was optimized using bioinformatics software to adapt to high-efficiency expression in Escherichia coli, and then the coding sequences were cloned into the expression papilloma In the recombinant plasmid of virus (PV) L1 protein, the insertion site can be DE loop, BC loop, FG loop or HI loop. Afterwards, the L1 recombinant plasmid of the chimeric SARS-CoV-2 spike protein epitope polypeptide coding sequence was transformed into Escherichia coli for expression.

[0118] Specifically: first use the online bioinformatics software CodonAdaptationTool (JCAT) (http: / / www.jcat.de / ) to optimize the coding SARS-CoV-2 spike protein epitope polypeptide SEQ ID NO: 1, SEQ ID NO: 2 and the codon sequence of SEQID NO:3. The codon-optimized gene sequence was outsourced and ...

Embodiment 2

[0120] Embodiment 2: Expression and purification of HPV L1 protein of chimeric SARS-CoV-2 spike protein epitope polypeptide

[0121] Identify the correct recombinant plasmid, and transform the recombinant plasmid of the chimeric SARS-CoV-2 spike protein epitope polypeptide into the Escherichia coli expression strain BL21. Pick a single clone and inoculate it into LB liquid medium containing ampicillin resistance, culture overnight at 37°C, inoculate the culture in fresh LB medium containing ampicillin resistance at a volume ratio of 1:100, and culture it with shaking at 37°C until When OD600nm is about 0.8, add IPTG with a final concentration of 1 mM, induce expression at 37°C for 6 hours, or induce overnight at 25°C. The cells induced to express the fusion protein at 37°C were collected by centrifugation. Cells were lysed with a high-pressure disruptor and centrifuged at high speed (12,000g) for 1 hour. The supernatant was collected and 20% ammonium sulfate was added for pri...

Embodiment 3

[0122] Example 3: Identification of HPV L1-VLP of Chimeric SARS-CoV-2 Spike Protein Epitope Polypeptide

[0123] After completing all the purification steps, the HPV L1-VLP sample of the obtained chimeric SARS-CoV-2 spike protein epitope polypeptide was subjected to dynamic light scattering and electron microscope observation to identify its particle size and shape.

[0124] Specifically, after the HPV L1-VLP solution of the chimeric SARS-CoV-2 spike protein epitope polypeptide obtained after purification was filtered through a 0.22 μm filter membrane, the particle size distribution and particle uniformity of the particles were analyzed using a Malvern dynamic light scattering instrument. Once the situation. image 3 Shown is the HPV16 VLP particle size detection result of the chimeric novel coronavirus spike protein epitope polypeptide.

[0125] Z average particle size (d.nm): 51.48; PdI: 0.012; intercept: 0.961; product quality: good.

[0126] Table 1: Intensity-force-diam...

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Abstract

The present disclosure relates to, for example, a virus-like particle (VLP) composition assembled from a papillomavirus (PV) (e.g., HPV or BPV) L1 major capsid protein comprising chimeric polypeptideepitopes, and surface spike epitope polypeptide of SARS-CoV-2 is inserted into the PV L1 major capsid protein. The invention also relates to a vaccine composition comprising VLPs.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular to a novel coronavirus (SARS-COV-2) polypeptide vaccine using papillomavirus (PV) virus-like particles (VLP) to present antigens. Background technique [0002] The causative agent of COVID-19, SARS-COV-2, belongs to the family of coronaviruses (Coronaviruses, CoVs). Coronaviruses are a class of enveloped positive-sense single-stranded RNA viruses that can infect humans and animals. They can be further subdivided into four genera based on genotype and serological characteristics: alpha-, beta-, gamma-, and delta-coronaviruses. To date, all coronaviruses confirmed to infect humans belong to the first two genera, Alpha- and Beta-coronaviruses. Among them, Beta-coronavirus has attracted people's attention, because members of the Beta-coronavirus family have caused three pandemics in the past 20 years, including the outbreak of SARS (caused by SARS-CoV) in 2003, and MERS (caused b...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N1/21A61K39/215A61P31/14C12R1/19
CPCC07K14/005C12N15/70A61K39/12A61P31/14C12N2710/20023C12N2710/20022C12N2770/20022C07K2319/00C12N2770/20034A61K2039/5258
Inventor 蒋敦泉伍树明刘永江张海江张士彬陈晓王学红银飞
Owner BEIJING HEALTH GUARD BIOTECH
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