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Antibody for preventing or treating cancer

An antibody, cancer technology, applied in the direction of antibodies, applications, anti-tumor drugs, etc., can solve the problems of Claudin18.2 unable to play normal functions and the destruction of tight junctions

Active Publication Date: 2020-11-20
北京亦庄国际蛋白药物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In tumors, the tight junctions between cells are disrupted and Claudin18.2 cannot perform its normal function

Method used

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  • Antibody for preventing or treating cancer
  • Antibody for preventing or treating cancer
  • Antibody for preventing or treating cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Construction of HEK293 cells expressing Claudin18.1 and Claudin18.2 antigens

[0066] The pcDNA3.1 vector encoding human Claudin18.1 and Claudin18.2 antigen genes (purchased from Invitrogen) was transfected into HEK293 cells (purchased from ATCC), and 200 μg / mL geneticin was used as the selection pressure to obtain stable expression of Claudin18.1 and Claudin18.2 antigen HEK293 cells. The Claudin18.2 antibody IMAB362 of Ganymed Company was used as a positive antibody (self-made, CHO-S cell (Invitrogen) transient expression and one-step affinity chromatography purification, the specific method refers to Examples 2 and 3, that is, BY0-0 is positive Antibody), HEK293 cells stably expressing human Claudin18.1 and Claudin18.2 antigens were screened by FACS method.

Embodiment 2

[0067] Example 2 Construction of Candidate Antibody BY4-8 Expression Vector

[0068] The HindIII restriction site ( AAGCTT), KoZAK sequence ( GCCGCCACC ), ATG, signal peptide gene GAGAGAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGT and antibody heavy chain coding gene (including heavy chain variable region coding gene SEQ ID NO: 18 and constant region IgG1 coding gene SEQ ID NO: 20), termination code TAA and EcoRI coding gene GAATCC Sequential fusion in series, and the use of chemical synthesis to obtain gene fragments. Through the EcoRI and HindIII sites, the above fragment was inserted into the eukaryotic expression plasmid pCDNA 3.4(+) (purchased from Invitrogen) and verified by sequencing to obtain the expression plasmid pCDNA3.4(+)-BY4-7 of the antibody heavy chain.

[0069] The HindIII restriction site ( AAGCTT ), KoZAK sequence ( GCCGCCACC ), ATG, signal peptide gene GAGAGAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGT and antibody light chain coding ...

Embodiment 3

[0073] Example 3 Expression and purification of anti-Claudin18.2 antibody

[0074] Using the DNA constructs described in Example 2, they were respectively transiently transfected into CHO-S cells (purchased from Invitrogen) to express the target antibody. According to the CHO-S cell operation manual provided by the manufacturer (Freedom TM CHO-S TM KitUSER GUIDE), adjust the cell density to 1x10 the day before plasmid transfection 6 individual / ml. On the day of plasmid transfection, mix with the transfection reagent and add to EXPICHO EXPRESSION MEDIUM cell culture medium (purchased from Invitrogen Company), 37°C, 8% CO2 continued to culture until the 8th day, collected the cell liquid, and removed the cells by centrifugation, 0.2 After μm filtration, Protein A was purified by affinity chromatography, the pH of the collected samples was adjusted to 5.5, and stored at 2-8°C. The purified antibody was detected by SDS-PAGE and SEC-HPLC, and the purity was above 95%.

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Abstract

The invention belongs to the technical field of antibodies, and provides an antibody for preventing or treating cancer. The antibody specifically binds to human claudin 18.2. The invention also provides a pharmaceutical composition comprising the antibod, application of the composition, nucleic acid molecules encoding the antibody, a vector comprising the nucleic acid molecules, host cells and a method for preparing the antibody.

Description

technical field [0001] The invention relates to an antibody for preventing or treating cancer, especially advanced gastric cancer, and belongs to the field of biotechnology. Background technique [0002] Gastric cancer is one of the most common cancers worldwide and the second most common cancer in China. According to the disclosure at the 12th International Gastric Cancer Conference held in Beijing in April 2017, about 680,000 new cases of gastric cancer are discovered in China every year, accounting for about half of the total number of cases in the world. Compared with the 446,500 cases announced in 2012, the average annual increase rate exceeded 13%. The death rate of gastric cancer in China is 4 to 8 times that of developed countries in Europe and the United States. About every 2 to 3 minutes, one Chinese person dies of gastric cancer. Compared with other countries, the situation of gastric cancer in China is more severe. [0003] When most gastric cancer patients in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12N15/13A61K39/395A61P35/00B65D25/02
CPCC07K16/28A61P35/00B65D25/02C07K2317/92C07K2317/56C07K2317/565C07K2317/76C07K2317/94A61K2039/505
Inventor 梁爽刘珊侯建华
Owner 北京亦庄国际蛋白药物技术有限公司
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