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Polypeptide targeting novel coronavirus COVID-19 and application of polypeptide

A technology for COVID-19 and coronavirus, applied in the field of peptides targeting the new coronavirus COVID-19, can solve the difficult problems of effective treatment of coronavirus, and achieve the effect of enriching sequence diversity, ensuring efficiency and small molecular weight

Active Publication Date: 2020-08-07
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The target specificity and highly mutagenicity of coronaviruses make effective treatment of coronaviruses quite difficult

Method used

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  • Polypeptide targeting novel coronavirus COVID-19 and application of polypeptide
  • Polypeptide targeting novel coronavirus COVID-19 and application of polypeptide
  • Polypeptide targeting novel coronavirus COVID-19 and application of polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1 Biotin or FITC labeling of S protein

[0092] In this example, the S protein (SEQ ID NO: 29) is first labeled with biotin or FITC, and the steps are as follows:

[0093] 0.15 mol / L NaCl solution and 0.15 mol / L NaHCO at pH=9.0 3 -Na2 CO 3 The buffer solution was mixed at a molar ratio of 9:1, and protein S was added to the mixture for protein dissolution, so that the final concentration of protein S was 10 mg / mL;

[0094] Add biotin or FITC to the S protein solution, place in an ice bath, and magnetically stir overnight in the dark;

[0095] On the second day, the mixed solution was placed in a dialysis bag with a molecular weight cut-off of 200, dialyzed in deionized water for two days, and the water was changed several times;

[0096] Finally, the dialyzed solution was collected and freeze-dried by a freeze dryer to obtain biotin- or FITC-labeled S protein.

Embodiment 2

[0097] Embodiment 2 Establishment of OBOC polypeptide library

[0098] In this example, the OBOC method is used to synthesize a peptide library on resin beads (beads), and the steps are as follows:

[0099] (1) Take 1.8 g of resin beads and put them in the synthesis tube, add a certain amount of DMF to soak the resin for resin swelling, and soak for more than 2 hours;

[0100] (2) Add Fmoc-protected methionine (M) to the synthesis tube, and add a mixture of HOBT (1-hydroxybenzotriazole) and DIC (N,N'-diisopropylcarbodiimide) solution, reacted on a shaking table for more than 2 h; alternately rinsed with methanol and DMF three times, repeatedly blowing up the beads with a straw during the rinse process; then tested the beads in a ninhydrin boiling water bath, and the beads were colorless;

[0101] (3) Add a deprotection agent (20% hexahydropyridine + 80% DMF) for Fmoc deprotection for 10 min, alternately rinse with methanol and DMF three times, and repeatedly blow up the beads...

Embodiment 3

[0108] Example 3 Screening of polypeptides targeting S protein

[0109] The FITC-labeled S protein and the peptide library were incubated in PBS buffer for 2 hours, the luminescence was observed under a fluorescence microscope (excitation wavelength 405 nm), and the observed peptide resins with obvious green fluorescence were selected.

[0110] like figure 1 Shown is the fluorescent microscope picture of FITC-labeled S protein interacting with OBOC polypeptide library. The beads bound to S protein can be observed with green fluorescence, and the peptides that can bind to S protein are selected according to the fluorescence.

[0111] The selected polypeptide resin was washed several times with ultrapure water and dried, using 30% CNBr 3 The ethanol solution was lysed overnight, the supernatant was obtained by centrifugation, and the peptide sequencing was carried out after desalting treatment.

[0112] Exemplarily, Figure 2(A), Figure 2(B), Figure 2(C) and Figure 2(D) are the...

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Abstract

The invention provides a polypeptide targeting novel coronavirus COVID-19 and application of the polypeptide. The polypeptide comprises an amino acid sequence shown as SEQ ID NO: 1-28; or the polypeptide is a molecule which is obtained by adding 1-20 amino acid polypeptide sequences and / or conjugation connection carbon chains to the N end and / or the C end of SEQ ID NO: 1-28 and has the activity oftargeting coronavirus COVID-19. The polypeptide library is synthesized by adopting a combined chemical method of one bead and one compound (one-by-one-complex, OBOC), so that the polypeptide libraryis obtained. By utilizing the polypeptide library and the S protein on the surface of the novel coronavirus, the screened polypeptide has strong binding capacity with the S protein of the coronavirusCOVID-19, and a basis is provided for research and development of novel coronavirus detection reagents and treatment drugs.

Description

technical field [0001] The invention belongs to the field of biomedical technology, and relates to a polypeptide targeting novel coronavirus COVID-19 and its application. Background technique [0002] Virus is a type of microorganism with the smallest shape and the simplest structure. It belongs to non-cellular microorganisms. It has no typical cell structure and energy-generating enzyme system, and can only grow and reproduce in parasitic living cells. Viruses play a very important role in medical microbiology. Among the various diseases caused by microorganisms, viral diseases account for as high as 75%. Viral diseases are not only highly contagious and widespread, but also have few effective drugs. The large-scale coronavirus infectious pneumonia that has occurred in recent years has aroused widespread concern from all walks of life. The new type of coronavirus pneumonia has become the focus of attention and discussion. However, no specific drug has been developed for t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06G01N33/569A61K47/42A61P31/14
CPCA61K47/42A61P31/14C07K7/06G01N33/56983G01N2333/165G01N2469/10
Inventor 王浩王磊杨培培安红维赵宇亮
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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