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Novel coronavirus multi-epitope recombinant antigen and preparation method thereof

A technology for recombinant antigens and coronaviruses, applied in biochemical equipment and methods, viruses, viral peptides, etc., can solve the problems of being unable to be used in a large area by grassroots medical institutions, demanding requirements, and long detection time, so as to eliminate the risk of false positives, The effect of improving specificity and enhancing recognition ability

Pending Publication Date: 2020-05-26
杭州贤至生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method has high accuracy, it requires a special operating site, professional operators, and professional equipment. The detection time is too long and the requirements are relatively strict, so it cannot be used on a large scale in primary medical institutions.

Method used

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  • Novel coronavirus multi-epitope recombinant antigen and preparation method thereof
  • Novel coronavirus multi-epitope recombinant antigen and preparation method thereof
  • Novel coronavirus multi-epitope recombinant antigen and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Selection of novel coronavirus dominant antigenic epitopes

[0018] The biological software DNAstar was used to analyze the hydrophilicity, hydrophobicity and antigenicity of the epitope sequences of S, E, and M antigens of the novel coronavirus, and the dominant epitopes of the S antigen (SEQ ID No: 2) and the dominant epitopes of the E antigen (SEQ ID No: 2) were respectively selected. ID No:3) and M antigen dominant epitope (SEQ ID No:4). At the same time, the sequence comparison results showed that the selected dominant antigenic epitope had high sequence specificity and had no obvious homology with other antigenic sequences other than coronavirus antigens.

Embodiment 2

[0019] Example 2: S, E, M antigen dominant epitope tandem

[0020] In order to enhance the ability of the recombinant antigen to recognize the new coronavirus antibody, the dominant epitopes of the S, E, and M antigens were connected through a flexible fragment (GlyGlyGlyGlySer), and a His tag was added to the carbon-terminus of the sequence to obtain the amino acid sequence of the recombinant antigen. The specific sequence is as follows: The sequence listing is shown in SEQ ID No:1.

Embodiment 3

[0021] Example 3: Optimizing the Nucleotide Sequence Encoding the Recombinant Antigen

[0022] In order to increase the expression of recombinant antigen, under the premise that the amino acid sequence of the recombinant antigen remains unchanged, the amino acid sequence encoding the recombinant antigen is converted into the corresponding nucleotide sequence according to the preferred codons of CHO cells. The specific sequence is shown in the sequence table SEQ ID No: 5. shown, and the nucleotide sequences corresponding to the enzyme cutting sites EcoRI and BamHI were added to the upstream and downstream respectively, and synthesized by Hangzhou Xianzhi Biotechnology Co., Ltd. The synthesized target gene was cloned in the pMD19-T vector (Bao Bioengineering Dalian Co., Ltd.).

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Abstract

The invention belongs to the technical field of biology, and discloses a novel coronavirus multi-epitope recombinant antigen and a preparation method thereof. The recombinant antigen comprises a plurality of dominant antigen epitopes of the novel coronavirus, a CHO cell preferred codon is adopted to convert the amino acid sequence of the recombinant antigen into a corresponding nucleotide sequence, the nucleotide sequence is chemically synthesized, and a recombinant expression vector is constructed, so that the expression quantity of the recombinant antigen in the CHO cell is increased. In addition, it is verified that the recombinant antigen can generate immunological reaction with mouse serum obtained through immunization of different antigens of the novel coronavirus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and discloses a novel coronavirus multi-epitope recombinant antigen and a preparation method thereof. The invention relates to a recombinant antigen, which contains multiple dominant antigenic epitopes of the novel coronavirus, and adopts CHO cell preferred codons to convert the amino acid sequence of the recombinant antigen into a corresponding nucleotide sequence, and chemically synthesize the nucleotide sequence and construct a recombinant expression vector to increase the expression of the recombinant antigen in CHO cells. In addition, it has been verified that the recombinant antigen can produce an immunological reaction with mouse serum immunized with different antigens of the new coronavirus. Background technique [0002] The 2019 novel coronavirus, or "2019-nCoV", is a new strain of coronavirus that has never been found in humans before. The novel coronavirus is a new strain of coronavirus ...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/85C07K16/10C07K16/06G01N33/569G01N33/558
CPCC07K14/005C12N15/85C07K16/10C07K16/06C07K16/065G01N33/56983G01N33/558C12N2770/20022C07K2319/00C07K2319/21C12N2800/22C12N2800/107G01N2333/165G01N2469/20
Inventor 项美华王小娟曹丹琴武戌青冯俊涛余铭恩
Owner 杭州贤至生物科技有限公司
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