Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Neutralizing single-domain antibody with function of resisting novel coronavirus SARS-Cov-2 and application of neutralizing single-domain antibody

A sars-cov-2, single-domain antibody technology, applied in antiviral agents, antiviral immunoglobulins, applications, etc., can solve the problems of limited and difficult changes in the structure of S protein

Active Publication Date: 2020-12-11
NANJING MEDICAL UNIV
View PDF5 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limited structural changes of the S protein of SARS-CoV-2 caused by a single point mutation, it is difficult to obtain neutralizing antibodies against the mutant virus

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Neutralizing single-domain antibody with function of resisting novel coronavirus SARS-Cov-2 and application of neutralizing single-domain antibody
  • Neutralizing single-domain antibody with function of resisting novel coronavirus SARS-Cov-2 and application of neutralizing single-domain antibody
  • Neutralizing single-domain antibody with function of resisting novel coronavirus SARS-Cov-2 and application of neutralizing single-domain antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example is to screen all human single domain antibodies targeting SARS-CoV-2-RBD.

[0032] Tomlinson I&J phage library (Genservice Ltd, Cambridge, UK, the library size is 1.47×10) 8 ) antibody sequence as a template, clone the antibody heavy chain sequence of the library, and construct a single-domain antibody library. Using phage display technology, SARS-CoV-2-RBD-HIS (ARG330-VAL524) protein as antigen was screened in single domain antibody library.

[0033] The specific process is as follows:

[0034] The ELISA plate was coated with 50μg / ml SARS-CoV-2-RBD his antigen at 4℃ overnight, 50ul / well; Using PBS solution (PBS STM) containing 5% skim milk powder and 0.1% tween-20 to block the enzyme-labeled plate at room temperature for 1 hour; Single domain antibody phage library with 10 12 Pfu was mixed with 10% skim milk powder PBS solution (PBSM) 1:1, then incubated at room temperature. After 2 hours, it was transferred to a sealed ELISA plate of SARS-CoV-2-RBD his antig...

Embodiment 2

[0045] This example is the specific binding of phage 4D8 clone to SARS-CoV-2-RBD protein.

preparation Embodiment 1

[0046] The 4D8 phage of Example 1 was prepared, and the binding specificity of 4D8 clone with SARS-CoV-2-RBD protein was detected by Phage ELISA.

[0047] The specific process is:

[0048] The ELISA plate was coated with 5μg / ml of SARS-CoV-RBD-hFc, SARS-CoV-2-RBD-his and SARS-CoV-2-RBD-hFc at 4℃ overnight. Seal that enzyme labeled plate with 3% pbstm at room temperature for 1 hour; 4D8 phage was incubated with 6% PBSM at the ratio of 1:1 at room temperature for 2 hours, then added to the sealed enzyme-labeled plate (50μl / well), and incubated at room temperature for 1 hour; The enzyme-labeled plate was washed with 0.05％PBST for 3 times (340ul / well); The HRP / anti-M13 monoclonal conjugate was mixed with 5% pbstm at the ratio of 1: 4000, added into the washed enzyme-labeled plate (50μl / well), and incubated at room temperature for 1 hour; Wash the enzyme-labeled plate with 0.05％PBST for 5 times; Add TMB chromogenic solution into enzyme-labeled plate (100μl / well), develop color at r...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a neutralizing single-domain antibody with a function of resisting novel coronavirus SARSCov2 and application of the neutralizing single-domain antibody. The antibody has at least one of a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3. The antibody can be used for preparing a diagnostic reagent or a diagnostic kit, an antibody medicine or a medicinal composition against COVID-19. An SARS-CoV-2 specific human monoclonal single-domain antibody is obtained by a phage display technology, has an SARS-CoV-2 pseudo-virus neutralizing effect, and has a synergistic neutralizing effect on D614G mutant strain pseudovirus when being combined with an IgG type SARS-CoV-2 neutralizing antibody 4A3 for use. Therefore, the antibody provides an efficient alternative antibody medicine for prevention and treatment of COVID-19.

Description

Technical field [0001] The invention relates to a neutralizing single-domain antibody against novel coronavirus SARS-Cov-2 and its application, belonging to the technical field of biomedicine. technical background [0002] The outbreak of novel coronavirus (COVID-19) at the end of 2019 is an acute respiratory infectious disease caused by novel coronavirus (SARS-CoV-2) infection. [1] . SARS-CoV-2 is a positive stranded RNA virus with a diameter of about 80-120nm, and it is a β coronavirus. [2] , which is very similar to the structure of SARS-CoV virus, and its genome homology is as high as 79.5%. [3] . Studies have confirmed that both SARS-Cov-2 and SARS-CoV host receptors are angiotensin converting enzyme 2(ACE2). [3] . ACE2 is a carboxypeptidase and a protein receptor, which is mainly expressed in alveolar cells and intestinal epithelial cells. [4] . SARS-CoV-2 binds to ACE2 through the receptor binding domain,RBD) of spike protein S1 subunit on the virus coat, which induces the...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/569A61K39/42A61P31/14
CPCA61K2039/505A61P31/14C07K16/10C07K2317/565C07K2317/569C07K2317/76C07K2317/92G01N33/56983G01N2333/165
Inventor 高威马素娟叶伟敖磊刘晓宇
Owner NANJING MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com