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

High-affinity anti-MSLN antibody and applications thereof

A technology of antibody and carrier, applied in the field of biomedicine, can solve the problems of high mortality rate, less than 6%, lack of

Pending Publication Date: 2018-03-27
SHANGHAI GENBASE BIOTECH CO LTD
View PDF5 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pancreatic cancer is a malignant tumor with a very high mortality rate. According to the statistics of the World Health Organization, there are 39,287 new cases of pancreatic cancer patients in China every year, and the one-year survival rate is less than 2%. So far, there is no effective therapy that can be applied to the treatment of pancreatic cancer; Carcinoma is a common malignant tumor that threatens the survival of women. There are 34,351 new cases of ovarian cancer patients in China every year, and the one-year survival rate is less than 6%. So far, there is no effective therapy for the treatment of ovarian cancer
Drugs approved by the FDA for pancreatic cancer mainly include tubulin inhibitors (Abraxane), PI3K inhibitors (Everolimus), EGFR inhibitors (Erlotinib) and multi-target inhibitors (Sunitinib), and the clinical benefits of these drugs are not high In addition, the FDA-approved drugs for ovarian cancer mainly include VEGFR antibody (Bevacizumab), platinum (Cisplatin), cyclophosphamide, tubulin inhibitors, etc., and their clinical benefits are also low

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
  • High-affinity anti-MSLN antibody and applications thereof
  • High-affinity anti-MSLN antibody and applications thereof
  • High-affinity anti-MSLN antibody and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0192] Preparation of monoclonal antibodies

[0193] Antibodies of the present invention can be prepared by various techniques known to those skilled in the art. For example, an antigen of the invention may be administered to an animal to induce the production of monoclonal antibodies. For monoclonal antibodies, hybridoma technology can be used to prepare (see Kohler et al., Nature 256; 495, 1975; Kohler et al., Eur.J.Immunol.6:511, 1976; Kohler et al., Eur.J.Immunol. 6:292,1976; Hammerling et al., In Monoclonal Antibodies and T Cell Hybridomas, Elsevier, N.Y., 1981) or can be prepared by recombinant DNA methods (US Patent No. 4,816,567).

[0194] Representative myeloma cells are those that fuse efficiently, support stable high-level production of antibody by selected antibody-producing cells, and are sensitive to culture medium (HAT medium matrix), including myeloma cell lines, such as murine Myeloma cell lines, including those derived from MOPC-21 and MPC-11 mouse tumors (...

Embodiment 1

[0226] Example 1. Construction of CDR1, CDR2, and CDR 3 mutation libraries of heavy chain (H) and light chain (L) of MN single-chain antibody

[0227] Using the pCAN-scFv MN plasmid (purchased from GE) as a template, mutations were introduced by PCR with random primers, and the primers are shown in Table 1. The obtained PCR products of the heavy chain (H) and light chain (L) of the scFv MN CDR1, CDR2, and CDR3 mutation library were named H1, H2, H3, L1, L2, and L3, respectively. After the PCR product and pCANTAB 5E were digested and recovered with Sfi I and Not I, they were ligated overnight at 16°C with T4 DNA ligase. The ligation product was electroporated to TG1, resuspended in 2YT and recovered at 37°C for 1 hour, then the bacterial solution was taken for gradient dilution and plate counting, and the storage capacity of each mutant library was at least 108, and the rest of the bacterial solution was all coated with 2YT (GA: 5% glucose, 100ug / ml penicillin) plate. 20 sin...

Embodiment 2

[0230] Example 2. Panning of phage antibody library

[0231] Add 20nM MSLN-Fc-biotin antigen (MSLN was purchased from SinoBiological; Biotin labeling reagent was purchased from Sigma) and incubate with the phage antibody library at room temperature for 2h, then transfer the mixture to streptavidin magnetic beads (purchased from LifeTechnology) and incubate at room temperature 15min. Unbound phages were washed away with PBST-PBS, and trypsin was added for 30 min at 37°C to elute bound phages. Infect 4ml of TG1 cells in the logarithmic phase with the phages digested and eluted by trypsin, let stand at 37°C for 30 minutes, take part of the bacterial liquid to serially dilute for plate counting, and coat the rest of the bacterial liquid on 2YT (GA) plates for next A round of packaging. The packaged phages can be used for the next round of panning. A total of 4 rounds of panning and enrichment were carried out. In each round of panning, 10-fold dilution gradients decreased the an...

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
concentrationaaaaaaaaaa
Login to View More

Abstract

The present invention provides a high-affinity anti-MSLN antibody and applications thereof, and specifically relates to an antibody having high affinity to MSLN, wherein the antibody is a single-chainantibody, has a heavy chain variable region and a light chain variable region, can be used for anti-MSLN-based chimeric antigen receptor T cell therapy methods, and has significant antitumor effects.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular, the invention relates to a high-affinity anti-MSLN antibody and its application. Background technique [0002] MSLN was highly expressed in ovarian cancer, mesothelioma and pancreatic cancer, among which 66% of ovarian cancer samples (10 / 15) MSLN highly expressed; 100% of pancreatic cancer samples (4 / 4) MSLN highly expressed. Pancreatic cancer is a malignant tumor with a very high mortality rate. According to the statistics of the World Health Organization, there are 39,287 new cases of pancreatic cancer patients in China every year, and the one-year survival rate is less than 2%. So far, there is no effective therapy that can be applied to the treatment of pancreatic cancer; Cancer is a common malignant tumor that threatens the survival of women. There are 34,351 new cases of ovarian cancer in China every year, and the one-year survival rate is less than 6%. So far, there is no effectiv...

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): C07K16/30C07K16/28C07K19/00C12N15/13C12N15/62C12N5/10C12N15/85G01N33/68G01N33/574G01N33/577A61K39/395A61P35/00
CPCC12N5/0636C07K16/28C07K16/30C12N2800/107A61K2039/505A61K2039/545C07K2319/21C07K2317/56C07K2317/565C07K2317/24
Inventor 牟男张云马泽龙袁纪军曹跃琼
Owner SHANGHAI GENBASE BIOTECH CO LTD
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