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

Antibody or antigen-binding fragment thereof that specifically recognizes b cell malignancies, chimeric antigen receptor comprising same, and uses thereof

A technology that combines fragments and antigens, applied in receptor/cell surface antigen/cell surface determinant, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, animal cells, etc. Cancer deaths and more

Pending Publication Date: 2020-08-14
ABCLON
View PDF11 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some patients who do not respond to available treatments or who relapse, which is the leading cause of death from childhood cancer

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
  • Antibody or antigen-binding fragment thereof that specifically recognizes b cell malignancies, chimeric antigen receptor comprising same, and uses thereof
  • Antibody or antigen-binding fragment thereof that specifically recognizes b cell malignancies, chimeric antigen receptor comprising same, and uses thereof
  • Antibody or antigen-binding fragment thereof that specifically recognizes b cell malignancies, chimeric antigen receptor comprising same, and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0201] Example 1: Development of antibodies related to CD19

[0202] In order to develop antibodies, the extracellular domain (ECD, Extracellular domain) of human CD19 protein is produced in animal cells and used as an antigen. Bind the C-terminus of the extracellular structure to the hinge of human immunoglobulin G1 (IgG1) and the Fc site (CH 2 -CH 3 ) morphological deoxyribonucleic acid (DNA) cloned in pCEP4 vector (Invitrogen, USA). Subsequently, the cloned above vectors were transiently transformed into FreeStyle TM 293F (Invitrogen, USA) cells to obtain CD19-ECDFc fusion protein. Phage bio-panning was performed by using the OPLA library with the CD19-ECDFc fusion protein described above. The antibody library uses VCSM13 helper phage (helper phage) to obtain the antibody library in the form of phage and use it for panning. The number of library phages that initially bind to the antigen is 10 13 . The panning rounds (panning round) were carried out to 4 rounds, and as...

Embodiment 2

[0206] Example 2. Enhanced affinity for developed antibody fragments

[0207] In order to obtain antibody fragments that bind CD19 better than CD19_8.1, a new sub-library was prepared by combining the heavy and light chain libraries. To generate sub-libraries, oligonucleotides containing NNK degenerate codons were used. CD19_8.1 antibody fragment was used as template DNA. Random codons were introduced into five complementarity determining regions (CDRs) except HCDR3 by polymerase chain reaction (PCR). The amplified antibody fragments were purified with QIAquick Gel Extraction Kit (QIAGEN, USA). The antibody fragment and the pComb3XSS vector were cut with sfiI restriction enzyme, and after ligation, they were transfected into ER2537 to prepare a phage library. Based on the prepared phage library, antibodies were screened in the same manner as in Example 1.

[0208] In order to screen antibodies with higher affinity among the selected antibodies, an enzyme-linked immunosorbe...

Embodiment 3

[0219] Example 3: Preparation of lentiviruses comprising chimeric antigen receptors linked to developed antibody fragments

[0220]A chimeric antigen receptor was developed using the developed antibody CD19_8.1. After codon-optimizing the CD8 leader sequence, CD19_8.1 in the form of single-chain variable region (scFv), CD8 hinge region, CD137 transmembrane region, and CD3ζ cytoplasmic region, the pLenti6-V5 / DEST was slowed down by SpeI / PacI Viral vectors (Invitrogen, USA) cut and ligated chimeric antigen receptors. The prepared construct (sequence No. 181 in the sequence listing) was confirmed by base sequence analysis.

[0221] The prepared lentiviral construct was transduced into the Lenti-X 293T (Takara BioInc, Japan) cell line together with pCMV-dR8.91, which is a vesicle containing a protein encoding as a viral capsid protein. Stomatitis Indiana virus G protein (VSV-G, vesicular stomatitis indiana virus G protein) nucleic acid and gag, pol and rev gene plasmids. Transd...

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

The present invention relates to: a novel antibody or an antigen-binding fragment thereof for use in the treatment of cancer by targeting B cell malignancies; a chimeric antigen receptor comprising the same; and uses thereof. The antibody of the present invention is an antibody specifically binding to CD19 highly expressed in cancer cells (particularly, blood cancer) and has very low homology compared to the CDR sequences of conventional CD19 target antibodies, and thus the sequence thereof is unique. In addition, cells expressing a chimeric antigen receptor comprising an anti-CD19 antibody orantigen-binding fragment of the present invention induce immune cell activity in response to a positive cell line expressing CD19, and thus can be usefully used as a therapeutic agent for CAR-immunecells.

Description

technical field [0001] This invention was completed with the support of multiple departments in South Korea through the project number 9991006240. The research and management agency for the above-mentioned topic is the new drug development business group of each department (financial), and the name of the research business is "full-cycle new drug development by each department". The name is "Development of a novel CD19 antibody-based CAR-T therapeutic agent", the competent authority is "abclon (strain)", and the research period is from February 01, 2018 to January 31, 2019. [0002] This patent application claims priority from Korean Patent Application No. 10-2017-0178559 filed with the Korean Patent Office on December 22, 2017, the disclosure of which is incorporated herein by reference. [0003] The present invention relates to novel antibodies or antigen-binding fragments thereof for use in treating cancer by targeting malignant B cells, chimeric antigen receptors comprisin...

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): C07K16/28C07K14/725C12N5/0783A61K35/17
CPCC07K16/2809C07K14/7051C07K2319/03C07K2317/622C07K2317/21C07K2317/92C07K2317/565C07K2317/73C12N5/0636A61K39/4611A61K39/4631A61K39/464412C07K16/2896C07K2317/56C07K2319/00C12N2510/00A61K39/4612A61P35/02A61P35/04A61K2121/00A61K2300/00C07K16/2803
Inventor 李种瑞金奎兑高凤国金起贤
Owner ABCLON
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