A method for expression and activity determination of Anti-Her2 humanized monoclonal antibody
10.3969/j.issn.1001-1978.2021.06.025
- Author:
Guan-Ping CHEN
1
;
Yi-Fan WANG
1
;
Xu-Hua YING
1
Author Information
1. Cancer Institute of Integrative Medicine, Tongde Hospital of Zhejiang Province
- Publication Type:Journal Article
- Keywords:
CHO cells;
HER2;
humanized;
monoclonal antibody;
purification;
recombinant
- From:
Chinese Pharmacological Bulletin
2021;37(6):884-888
- CountryChina
- Language:Chinese
-
Abstract:
Aim To establish a method for the stable expression of recombinant humanized Anti-her2 monoclonal antibody and to measure its bioactivity. Methods Anti-her2 heavy chain and light chain eukaryotic expression vectors were used to transfect CHO cells, and then scaled-up after screening the stably expressed cell line. Protein A affinity chromatography was utilized to purify the fusion protein. Western blot was used to identify the properties of the product, SDS-PAGE and HPLC were employed to detect its specificity, and CCK8 assay was used to detect its effect on cell viability. Results One cell line with stable expression of recombinant antibody was selected for scale-up. The supernatant was gained for the purification with protein concentration of 135.9 mg · L
