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New coronavirus vaccine based on chimpanzee adenovirus type 68 and MERS-CoV full length membrane protein

An adenovirus and chimpanzee technology, applied in the field of new coronavirus vaccines, has achieved wide application prospects, easy production and storage, and high titer effects

Active Publication Date: 2019-12-27
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are no specific drugs and vaccines for the treatment and prevention of this disease

Method used

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  • New coronavirus vaccine based on chimpanzee adenovirus type 68 and MERS-CoV full length membrane protein
  • New coronavirus vaccine based on chimpanzee adenovirus type 68 and MERS-CoV full length membrane protein
  • New coronavirus vaccine based on chimpanzee adenovirus type 68 and MERS-CoV full length membrane protein

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, the preparation of recombinant virus

[0047] 1. Construction of recombinant plasmids

[0048] 1. Insert the DNA molecule shown in Sequence 2 of the sequence listing between the XbaI and NheI restriction sites of the pShuttle-CMV vector to obtain the recombinant plasmid pShuttle-MERS-CoV Spike.

[0049] The DNA molecule shown in Sequence 2 of the Sequence Listing encodes the protein shown in Sequence 1 of the Sequence Listing. The protein shown in sequence 1 of the sequence listing is the full-length MERS-CoV Spike protein.

[0050] 2. Digest the recombinant plasmid pShuttle-MERS-CoV Spike with endonucleases PI-SceI and I-CeuI, and recover a DNA fragment of about 5019 bp.

[0051]After sequencing, the DNA fragment is shown as sequence 3 in the sequence listing. In sequence 3 of the sequence listing, nucleotides 238-617 are the CMV enhancer, nucleotides 618-821 are the CMV promoter, and nucleotides 937-4998 are the coding region of the full-length MERS-C...

Embodiment 2

[0088] Embodiment 2, the application of recombinant virus

[0089] 1. Animal immunity

[0090] 6-week-old female BALB / C mice were divided into 12 groups, 5 mice in each group, and were treated as follows:

[0091] The first group (G1 group): a single immunization was carried out by nasal drops, and the immunization substance was PBS buffer solution with pH 7.2;

[0092] The second group (G2 group): a single immunization was carried out by intramuscular injection, and the immune substance was PBS buffer solution with pH 7.2;

[0093] The third group (G3 group): a single immunization was carried out by nasal drops, and the immune substance was AdC68 virus liquid (the virus amount was 2×10 8 vp);

[0094] The fourth group (G4 group): a single immunization was carried out by nasal drops, and the immune substance was AdC68 virus liquid (the virus amount was 2×10 10 vp);

[0095] The fifth group (G5 group): a single immunization was carried out by intramuscular injection, and t...

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Abstract

The present invention discloses a new coronavirus vaccine based on chimpanzee adenovirus type 68 and MERS-CoV full length membrane protein. The recombinant adenovirus is obtained by transfecting an adenovirus packaging cell with a recombinant plasmid and then performing cell culture; the recombinant plasmid is obtained by inserting a specific DNA molecule into a delta-E1 region of a chimpanzee adenovirus vector AdC68; the specific DNA molecule has a full-length MERS-CoV Spike protein encoding gene; and the adenovirus packaging cell has an adenovirus E1 gene. The present invention also protectsthe recombinant adenovirus expressing the full-length MERS-CoV Spike protein; and a starting strain of the recombinant adenovirus is chimpanzee adenovirus type 68 or non-replicating chimpanzee adenovirus type 68. The developed vaccine against the new coronavirus MERS-CoV has important theoretical guidance value and broad application prospects, and provides a possibility for radical cure of MiddleEast respiratory syndrome.

Description

technical field [0001] The present invention relates to a novel coronavirus vaccine based on chimpanzee adenovirus type 68 and MERS-CoV full-length membrane protein. Background technique [0002] The new type of coronavirus (MERS-CoV) was first found to infect humans in the Middle East in 2012, and then this virus infection disease appeared in several European countries and regions. Most patients with MERS-CoV infection will develop severe respiratory disease, and its clinical symptoms are very similar to the disease caused by SARS-CoV that broke out in 2003. Because the disease can be transmitted from person to person, it has attracted great attention around the world. So far, there are no specific drugs and vaccines for the treatment and prevention of this disease. [0003] MERS-CoV uses the membrane protein S (also known as S protein) on its surface to enter susceptible cells. The S protein consists of the following three domains: the S1 domain at the N-terminus, the S...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/861C12N15/50C07K14/165A61K39/215A61P31/14C12R1/93
CPCA61K39/12A61P31/14C07K14/005C12N7/00C12N15/86C12N2710/10021C12N2710/10043C12N2770/20022
Inventor 张林琦周东明贾雯旭张超史宣玲
Owner TSINGHUA UNIV
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