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

A group of PD-L1 monoclonal antibodies and their medical use

A PD-L1, monoclonal antibody technology, applied in the field of tumor and immunology drugs, can solve the problem of low tumor inhibition effect, and achieve the effect of promoting the secretion of cytokine IL-2 by T cells

Active Publication Date: 2021-05-28
DONGDA BIOSCIENCE INC (SUZHOU)
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although PD-L1 antibodies have low side effects in clinical trials, their tumor suppressive effect is often lower than that of PD-1 antibodies

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
  • A group of PD-L1 monoclonal antibodies and their medical use
  • A group of PD-L1 monoclonal antibodies and their medical use
  • A group of PD-L1 monoclonal antibodies and their medical use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 - Construction of mouse hybridoma antibody

[0037] The construction of mouse hybridoma antibody includes the following 6 steps.

[0038] (1) Mouse immunization and fusion of hybridoma cells. Human PD-L1-ECD-mFc fusion protein was used as antigen, fully emulsified with an equal volume of complete Freund's adjuvant (Sigma, Cat No: F5581), and subcutaneously immunized 6-8 week-old Balb / c mice (purchased from Zhaoyan (Suzhou) New Drug Research Center Co., Ltd.), the antigen immunization dose was 20 μg / monkey. Subsequently, mice were subcutaneously immunized three times with the same dose of antigen fully emulsified with incomplete Freund's adjuvant (Sigma, Cat No: F5506) every 2 weeks. After three immunizations, the serum titer of the mice was determined, and a booster immunization was carried out through the abdominal cavity 3 days before the fusion. Using PEG Hybri-Max (Sigma, CatNo:7181) as a fusion agent, mouse spleen cells and SP2 / 0 cells were mixed in a ...

Embodiment 2

[0057] Example 2 - Cloning of PD-L1 Antibody Variable Region Gene

[0058] The PD-L1 monoclonal hybridoma cell line was lysed with TRIzon (Cwbiotech, Cat No: CW0580), and the total RNA of the hybridoma cells was extracted. The RNA of hybridoma cells was reverse-transcribed into cDNA using HiFi Script cDNA Synthesis Kit (Cwbiotech, Cat No: CW2569). Use cDNA as a template and use degenerate primers to amplify the heavy chain of the antibody by PCR method (Kettleborough et al. (1993) Eur J Immunology 23:206-211; Strebe et al. (2010) Antibody Engineering 1:3-14) and light chain variable region genes. After the PCR amplification product was connected to the T / A carrier, the DH5a competent cells were transformed, plated and cultured overnight at 37°C. Pick a single clone from the culture plate, expand the culture, extract the plasmid, and determine the gene sequence of the antibody. According to the gene sequence of the antibody, its complementary determinants (CDR) and framework...

Embodiment 3

[0062] Example 3 - Humanization of murine PD-L1 antibodies 6C8D8, 6F9B1A6 and 10E9C7G2

[0063] (1) The humanization of PD-L1 antibody was carried out by the complementary determinant grafting method. First, the human germline antibody sequences with the highest homology to the light and heavy chain variable regions of the murine 6C8D8, 6F9B1A6 and 10E9C7G2 antibodies were searched in the IMGT database. The germline selected for the humanization of the light chain variable region of the 6C8D8 antibody is IGKV6-21*02, and the humanization of the heavy chain variable region is IGHV2-5*01. The germline selected for the humanization of the light chain variable region of the 6F9B1A6 antibody For IGKV1-9*01, IGHV1-69*08 was selected for the humanization of the heavy chain variable region. The germline selected for the humanization of the light chain variable region of the 10E9C7G2 antibody was IGKV1-39*01, and the heavy chain variable region was humanized Select IGHV6-1*01.

[006...

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

No PUM Login to View More

Abstract

A panel of PD‑L1 monoclonal antibodies and their medicinal uses. The invention belongs to the field of tumor therapy and molecular immunology; specifically, it relates to a group of anti-PD‑L1 monoclonal antibodies and their medical applications. The present invention obtains a group of anti-PD-L1 monoclonal antibodies with excellent effects in blocking the interaction between PD-L1 and PD-1 through hybridoma technology, and successfully performs humanization and affinity maturation on them . The antibody shows great application prospects in the preparation of related drugs for blocking and regulating the effect and level of PD-L1 and significantly enhancing the body's immunity, especially for the treatment of cancer-related drugs.

Description

technical field [0001] The invention belongs to the field of tumor and immunology drugs, and specifically, the invention relates to a group of anti-PD-L1 monoclonal antibodies and their medical applications. [0002] technical background [0003] Programmed death ligand PD-L1, also known as B7-H1 or CD274, is a type I transmembrane glycoprotein encoded by the CD274 gene and belongs to the B7 family with a molecular weight of about 40KDa. PD-L1 is composed of an IgV-like domain, an IgC-like domain, a transmembrane region, and a short cytoplasmic peptide (Keir et al., (2008) Annu RevImmunol 26:677-704; Lin et al. al., (2008) PNAS 105:3011-3016). PD-L1 is widely expressed in vivo, not only expressed in B cells, T cells, bone marrow cells and dendritic cells, but also expressed at low levels in some non-lymphoid organs, such as heart, lung, etc. (Yamazaki et al., (2002 ) Journal of Immunology 169:5538–45; Eppihimer et al., (2002) Microcirculation 9:133–45; Liu J et al., (2007) ...

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 Patents(China)
IPC IPC(8): C07K16/28C12N15/13A61K39/395A61K47/55A61P35/00
CPCA61K47/55A61P35/00C07K16/2827C07K2317/24C07K2317/56C07K2317/565C07K2317/76C07K2317/92
Inventor 周漫潘剑锋孙锴王振生陈俊勇孙健区日山
Owner DONGDA BIOSCIENCE INC (SUZHOU)
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