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Polypeptide for preventing novel coronavirus pneumonia COVID-19 and application thereof

A SEQIDNO.1-SEQIDNO.11, amino acid technology, which is applied in the fields of immunology, biotechnology and biomedicine, can solve the problem that the vaccine under development has not been approved for marketing, and can quickly respond to virus mutation, reduce research and development costs, and reduce Effects of antigenic loss

Pending Publication Date: 2022-02-18
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, all vaccines under development have not yet been approved for marketing

Method used

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  • Polypeptide for preventing novel coronavirus pneumonia COVID-19 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Selection method of candidate polypeptide sequences and method of synthesizing polypeptides.

[0033] 1. Selection of candidate polypeptides.

[0034] The functional protein homology between SARS-CoV-2 and SARS-CoV is very high. Based on the discovered SARS-CoV antigen and antigenic determinant spectrum, the parasite is replaced with the SARS-CoV-2 polypeptide sequence, and based on the SARS-CoV antigen A series of candidate polypeptides were designed by appropriately extending the position of the determinant and increasing the antigenic determinant.

[0035]Based on the RBD / ACE2 binding characteristics of SARS-CoV-2, a series of candidate polypeptides were designed, the amino acid sequences of which are shown in SEQ ID NO.1-SEQ NO.11.

[0036] 2. Polypeptide synthesis.

[0037] 1. The polypeptide whose amino acid sequence is shown in SEQ ID NO.1-SEQNO.11 was prepared by using the conventional method of solid-phase peptide synthesis, and the specific steps a...

Embodiment 2

[0052] Example 2 Experiments on the specific antigenicity of polypeptides shown in SEQ ID NO.1-SEQ NO.11.

[0053] The polypeptides obtained in Example 1 were reacted with 16 sera of new crown patients respectively. Since the new coronavirus-infected human serum contains anti-new coronavirus antibodies, and can react with the polypeptide antigen from the new coronavirus protein sequence, it is amplified by enzyme-linked immunoreaction and the color is positive. The specific steps are as follows.

[0054] 1) Coating: coating solution is 0.07M NaHCO 3 , 0.03M Na 2 CO 3 (pH=9.6), coating concentration 10 μg / well, overnight at 4°C.

[0055] 2) Washing: wash with PBST (pH=7.4, containing 0.05% Tween 20) 5 times, and pat dry.

[0056] 3) Blocking: the blocking solution is 1-3% BSA, 100 μL / well, at 37° C. for 2 hours.

[0057] 4) Washing: wash with PBST (pH=7.4, containing 0.05% Tween 20) 5 times, and pat dry.

[0058] 5) Add the primary antibody: the patient's antiserum was d...

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Abstract

The invention relates to the fields of immunological technology, biotechnology and biological medicine, in particular to polypeptide for preventing novel coronavirus pneumonia COVID-19 and application of the polypeptide. The amino acid sequence of the antigenic polypeptide is selected from any one of the following: 1) polypeptide with the amino acid sequence as shown in SEQ ID NO.1-SEQ ID NO.11; 2) polypeptide having 85% or more of the same amino acid sequence as the amino acid sequence as shown in SEQ ID NO.1-SEQ ID NO.11; and 3) polypeptide obtained by subjecting the amino acid sequences shown as SEQ ID NO.1-SEQ ID NO.11 to substitution and / or deletion and / or addition of one or more amino acid residues and having the same functions. The polypeptide for preventing novel coronavirus pneumonia COVID-19 provided by the invention is obtained by chemical synthesis of a specific antigenic fragment screened from a novel coronavirus S protein sequence, and an immunogen experiment proves that the polypeptide has relatively strong antigenicity, can induce a neutralizing antibody in a human body, and is expected to become a polypeptide vaccine antigen, and a new vaccine is provided for preventing novel coronavirus pneumonia COVID-19.

Description

technical field [0001] The invention relates to the fields of immunology technology, biotechnology and biomedicine, in particular to a polypeptide for preventing novel coronavirus pneumonia COVID-19 and its application. Background technique [0002] Coronavirus is a positive enveloped virus with RNA, and its genome size is about 26-32kb, which is the largest RNA virus known genome. Genomic RNA and phosphorylated nucleocapsid (N) proteins are buried in the phospholipid bilayer and covered by the spike glycoprotein trimer (S), the membrane (M) protein (type III transmembrane glycoprotein) and the envelope ( E) The protein is located between the S proteins of the viral envelope. Coronaviruses have a variety of hosts, including birds and mammals, especially bats. Coronavirus is a kind of virus that widely exists in nature and can cause multi-system diseases including respiratory tract, digestive tract and nervous system. Highly pathogenic coronavirus infection has become a pub...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165G01N33/569A61K39/215A61P31/14
CPCC07K14/005G01N33/56983A61K39/12A61P31/14C12N2770/20022G01N2333/165C12N2770/20034
Inventor 刘刚于文俊
Owner TSINGHUA UNIV
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