Polypeptide and application thereof to novel coronavirus detection and antibody or vaccine screening

A coronavirus and virus technology, applied in the field of biomedicine, can solve the problems of long-term human existence, long hidden time, and three-time transmission of the new type of pneumonia virus

Pending Publication Date: 2020-09-11
ACAD OF MILITARY SCI ACAD OF MILITARY MEDICAL SCI BEIJING INST OF LIFEOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still no effective treatment for this new type of virus, and it is highly infectious, spreads widely, and hides for a long time. A small number of virus-infected patients have no symptoms or mild symptoms. induce secondary or even tertiary transmission
In addition, if no effective vaccine is developed, the new type of pneumonia virus may accompany humans for a long time

Method used

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  • Polypeptide and application thereof to novel coronavirus detection and antibody or vaccine screening
  • Polypeptide and application thereof to novel coronavirus detection and antibody or vaccine screening
  • Polypeptide and application thereof to novel coronavirus detection and antibody or vaccine screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The synthesis of embodiment 1 polypeptide

[0055] Design the polypeptide sequence according to Table 1, from the C-terminal to the N-terminal of the sequence, the synthesis steps are as follows:

[0056] 1) Weigh 0.3g of resin into a glass reactor, add DCM (dichloromethane) to swell for 30 minutes;

[0057] 2) Take out the DCM, add 0.6g of the first amino acid in the amino acid sequence, add 0.6g of DIEA (diisopropylethylamine), DMF (dimethylformamide), DCM, react by bubbling nitrogen for 60 minutes, and take out The reaction solution was washed three times by adding DMF and MEOH;

[0058] 3) Add 0.6 g of the second amino acid in the amino acid sequence to the reactor, add HBTH (1-hydroxy-benzotrichlorazole tetramethylhexafluorophosphate) and DIEA, react by bubbling nitrogen for 30 minutes, and wash off the liquid. Ninhydrin detection followed by capping with pyridine and acetic anhydride. Finally, wash, add an appropriate amount of decapping solution to remove the ...

Embodiment 2

[0064] The preparation of embodiment 2 polypeptide chips

[0065] Select the three-dimensional D-modified glass slide as the substrate of the polypeptide chip;

[0066] All the polypeptide fragments synthesized in Example 1 and the positive control were spotted on the surface of the slide by a microarray spotter.

Embodiment 3

[0067] Example 3 Polypeptide chip detects effectiveness test of SARS-CoV-2 protein antibody

[0068] Sample collection:

[0069] The samples came from Peking Union Medical College Hospital. According to the "Diagnosis and Treatment Plan for Pneumonia Infected by Novel Coronavirus" (trial version 7), patients with mild symptoms of COVID-19 were diagnosed. There were 15 early COVID-19 patients, 13 flu patients with similar symptoms and 12 non-influenza patients.

[0070] Screening of serum samples:

[0071] 1. Sealing: Add a fence to the chip, add 400ul / well 5% milk (0.5g milk powder + 10ml PBST) and seal at room temperature for 0.5h;

[0072] 2. Sample preparation: mix 4ul serum + 400ul 5% milk;

[0073] 3. Adding samples: add the diluted serum sample to the peptide chip prepared in implementation 2, 400ul / well, incubate at room temperature for 1h;

[0074] 4. Washing: wash 3 times with 0.05% PBST, 10 min each time;

[0075] 5. Add fluorescent dyes: add Donkey-anti-hIgG-53...

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Abstract

The invention provides a polypeptide capable of being applied to 2019 novel coronavirus (SARS-CoV-2) detection and antibody or vaccine drug screening. The polypeptide comprises at least 15 continuousamino acid sequences in an S protein sequence, an N protein sequence or an orf1ab protein sequence of an SARS-CoV-2 virus protein, and comprises any amino acid sequence as shown in SEQ ID No.1 to 25.

Description

technical field [0001] The invention belongs to the field of biomedical technology, and relates to a polypeptide and its application in detection of 2019 novel coronavirus (SARS-CoV-2), antibody or vaccine drug screening. Background technique [0002] Since December 2019, many cases of viral pneumonia have been found around the world, and the diagnosis results are all viral pneumonia. On February 11, 2020, the World Health Organization officially named the new coronavirus as the 2019 novel coronavirus (SARS-CoV-2), which is 82% similar to the SARS virus. However, there is still no effective treatment for this new type of virus, and it is highly infectious, spreads widely, and hides for a long time. A small number of virus-infected patients have no symptoms or mild symptoms. Induce secondary or even tertiary transmission. In addition, if no effective vaccine is developed, the new type of pneumonia virus may accompany humans for a long time. [0003] The SARS-CoV-2 virus pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C07K16/10G01N33/569A61K39/215A61P31/14
CPCC07K14/005C07K16/10G01N33/56983A61K39/12A61P31/14C12N2770/20022C12N2770/20034A61K2039/53G01N2333/165
Inventor 于晓波张晓梅王聃王红叶梁特代佳宇
Owner ACAD OF MILITARY SCI ACAD OF MILITARY MEDICAL SCI BEIJING INST OF LIFEOMICS
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