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

ANTIBODIES AGAINST SARS-CoV-1 OR SARS-CoV-2 AND USES THEREOF

A sars-cov-2 and antibody technology, applied in the direction of antibodies, medical preparations containing active ingredients, applications, etc., can solve problems such as limited clinical effects

Pending Publication Date: 2021-11-02
YANG SHENG TANG +1
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patients with pneumonia caused by SARS-CoV-2 infection should only be given general supportive treatment, oxygen therapy measures and antiviral treatment, such as α-interferon, lopinavir / ritonavir, chloroquine phosphate, etc. These measures bring limited clinical efficacy

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
  • ANTIBODIES AGAINST SARS-CoV-1 OR SARS-CoV-2 AND USES THEREOF
  • ANTIBODIES AGAINST SARS-CoV-1 OR SARS-CoV-2 AND USES THEREOF
  • ANTIBODIES AGAINST SARS-CoV-1 OR SARS-CoV-2 AND USES THEREOF

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0242] Example 1: Synthesis of anti-SARS-CoV-2 receptor binding region RBD gene and construction of expression vector

[0243] Referring to the full gene sequence of SARS-CoV2-2 (MN908947.3), the nucleotide sequence encoding the SARS-CoV2-2 receptor binding domain RBD protein (corresponding to amino acids 316-550 of the S protein), according to the human code Sub-preferences are optimized. And, the signal peptide coding sequence (the guide sequence of human B2M) is connected to the N-terminal of the optimized nucleic acid sequence, and the polyhistidine polypeptide (6×His) that is convenient for affinity chromatography purification is connected to the C-terminal, and the protein Named as RBD-His, its nucleotide sequence is shown in SEQ ID NO:22, and its amino acid sequence is shown in SEQ ID NO:23. NEBuilder HiFi DNA AssemblyMaster Mix (NEB Company) was used to connect the above nucleotide sequence to the expression vector EIRBsMie-C18hA2dtSCT (enzyme cutting site is AgeI / Bgl...

Embodiment 2

[0244] Example 2: Expression and purification of the RBD antigen of SARS-CoV-2

[0245] 1. Expression of RBD antigen of SARS-CoV-2

[0246] with 3x10 6 The density of the ExpiCHO cells in the appropriate amount of medium ExpiCHO TM Expression Medium (Thermo Scientific Company) was cultured in Erlenmeyer shaker flasks at 37°C, 8% CO 2 , and cultured in a constant temperature shaker with an appropriate speed for 24 hours, until the cell density reached 6x10 6 Density.

[0247] Use ExpiFectamine according to the kit instructions TM The CHO Transfection Kit (ThermoScientific Company) transfected the vector EIRBsMie-RBDhis obtained in Example 1 into ExpiCHO cells. After continuing to culture under the same conditions for 17-24h, add the feed and enhancer provided in the kit, and replace the cells to 32°C, 5% CO 2 , and continue culturing for 6 days in a constant temperature shaker with an appropriate rotation speed.

[0248] 2. Purification of the RBD antigen of SARS-CoV-2 ...

Embodiment 3

[0251] Example 3: Obtaining of mouse monoclonal antibody

[0252] 1. Mice Immunization

[0253] Standard in vivo immunization methods were used. For details, see Ed Harlow et al., "Antibodies A Laboratory Manual", Cold Spring Harbor Laboratory 1988. The brief procedure is as follows:

[0254] Use the SARS-CoV-2 RBD protein obtained in Example 2 to mix and emulsify in equal volumes with Freund's adjuvant. BALB / c female mice aged 6-8 weeks were immunized by multi-point subcutaneous injection in the bilateral groin, and a booster immunization was given every 2 weeks after the initial immunization. Serum antibody titers were measured by indirect ELISA, and fusion experiments were performed 4 weeks later.

[0255] The spleen was boosted 72 hours before the fusion of mouse spleen cells and mouse myeloma cells (SP2 / 0). The antigen for this immunization was the antigen without adjuvant, and the concentration was diluted to 1 mg / mL. Inject 50 μL of protein longitudinally along the s...

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
Equilibrium dissociation constantaaaaaaaaaa
Login to View More

Abstract

The present invention relates to the fields of immunology and molecular virology, in particular the diagnosis, prophylaxis and treatment of coronaviruses, such as SARS-CoV-2 and / or SARS-CoV-1. In particular, the present invention relates to monoclonal antibodies against coronavirus, as well as compositions (e.g., diagnostic and therapeutic agents) comprising the antibodies. In addition, the invention also relates to application of the antibody. The antibodies of the invention can be used for diagnosing, preventing and / or treating coronavirus infection and / or diseases caused by the infection.

Description

technical field [0001] The invention relates to the field of immunology and molecular virology, especially the field of diagnosis, prevention and treatment of coronavirus. In particular, the present invention relates to anti-coronavirus monoclonal antibodies, and compositions (eg, diagnostic and therapeutic agents) comprising such antibodies. Furthermore, the present invention also relates to the use of said antibody. The antibody of the present invention can be used for diagnosing, preventing and / or treating coronavirus infection and / or diseases caused by said infection. Background technique [0002] Coronavirus (coronavirus) infection can lead to respiratory diseases in humans. Mild coronavirus infection can cause flu-like symptoms, and severe infection can develop into severe viral pneumonia, threatening human life and health. Coronaviruses can infect humans and animals at the same time. If some animal-derived coronaviruses break through the host barrier and infect huma...

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
IPC IPC(8): C07K16/10C12N15/13C12P21/02A61K39/42A61K45/06A61P31/14G01N33/569G01N33/577
CPCC07K16/10A61K45/06A61P31/14G01N33/56983G01N33/577G01N33/54306G01N33/581C07K2317/24C07K2317/56C07K2317/565C07K2317/567C07K2317/52C07K2317/76C07K2317/92A61K2039/505G01N2333/165
Inventor 袁权巫洋涛熊华龙张天英王邵娟蒋艺超侯汪衡施洋张雅丽罗文新夏宁邵
Owner YANG SHENG TANG
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