Preparation and identification of humanized monoclonal antibody against periostin
10.3969/j.issn.1000-484X.2024.12.028
- VernacularTitle:抗periostin人源化单克隆抗体的制备及鉴定
- Author:
Xuejiao LI
1
;
Hang ZHAO
;
Shuo ZHANG
;
Huiying KANG
;
Yannan ZHOU
;
Shuang JIA
;
Xu LU
;
Hongli ZHAO
;
Yang HAI
Author Information
1. 辽宁何氏医学院基础医学院,沈阳 110163
- Keywords:
Periostin;
Humanized monoclonal antibody;
Retinal fibrosis
- From:
Chinese Journal of Immunology
2024;40(12):2628-2633
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare a humanized monoclonal antibody against periostin and establish a stable cell line.Meth-ods:Based on anti-periostin mouse monoclonal antibody developed by our laboratory,total RNA was extracted,and variable region sequences were obtained by RT-PCR amplification of VH and VL genes.The mouse antibody CDR region was transplanted into the human antibody framework receptor region,and the gene was subcloned into the expression vector PATX-GS2,and stably transfected into CHO cells.Monoclonal cell lines were obtained by MSX pressure screening and limited dilution.Results:VH and VL genes were amplified by RT-PCR,and the sequence of the light and heavy chain variable region were determined.Antibody humanization were successfully stablished by CDR transplantation method a murine antibody to a human framework,and a eukaryotic expression plasmid was constructed,which was transfected into CHO cells for expression,and human anti-periostin antibody was successfully obtained.ELISA and Western blot results showed that the humanized antibody had good anti-periostin activities and binding affinity.Conclu-sion:In this study,anti-periostin humanized monoclonal antibody has been successfully prepared,which can specifically bind to peri-ostin proteins in vivo and have biological activity,providing scientific data for the precise treatment of retinal fibrosis,tissue and organ fibrosis,and malignant tumors.
