中国免疫学杂志2024，Vol.40Issue(12)：2628-2633,6.DOI:10.3969/j.issn.1000-484X.2024.12.028
抗periostin人源化单克隆抗体的制备及鉴定
Preparation and identification of humanized monoclonal antibody against periostin
摘要
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.关键词
Periostin/人源化单克隆抗体/视网膜纤维化Key words
Periostin/Humanized monoclonal antibody/Retinal fibrosis分类
医药卫生引用本文复制引用
李雪娇,赵航,张朔,康慧颖,周艳楠,贾爽,路旭,赵洪礼,海洋..抗periostin人源化单克隆抗体的制备及鉴定[J].中国免疫学杂志,2024,40(12):2628-2633,6.基金项目
辽宁省民生科技计划项目(2021JH2/10300028). （2021JH2/10300028）
