靶向CD47单克隆抗体结构表征与生物学特性分析  

Structural Characterization and Biological Properties Analysis of A Monoclonal Antibody Targeting CD47

作　　者：吴丹青 张明玉 丁子芳 赵燕佳 彭缨[1] WU Danqing;ZHANG Mingyu;DING Zifang;ZHAO Yanjia;PENG Ying(Wuya College of Innovation,Shenyang Pharmaceutical University,Shenyang 110016,China;Changchun Jinsai Pharmaceutical Co.,Ltd.,Changchun 130012,China)

机构地区：[1]沈阳药科大学无涯创新学院,辽宁沈阳110016 [2]长春金赛药业有限责任公司,吉林长春130012

出　　处：《工业微生物》2024年第6期142-146,共5页Industrial Microbiology

摘　　要：文章通过转染中国仓鼠卵巢(CHO)细胞系制备抗CD47单克隆抗体,并对其进行综合表征,实现了对其结构验证和作用原理的深入探究。具体采用超高效液相色谱-串联质谱(UPLC-MS/MS)对单抗分子质量、氨基酸覆盖率、二硫键连接方式进行结构表征,同时通过阻断试验研究其与CD47的相互作用。结构确证显示,单抗分子质量与理论一致,氨基酸覆盖率达100%;二硫键配对方式与理论一致。阻断实验证明,anti-CD47可与CD47-his结合,并阻断CD47-his与信号调节蛋白α(SIRPα)的结合。该靶向单抗的分子结构与理论设计结构一致,可特异性结合CD47,阻断CD47-SIRPα信号轴,显示出了潜在的抗肿瘤效果。Anti-CD47 monoclonal antibodies were expressed by transfecting Chinese hamster ovary(CHO)cell lines,and a comprehensive characterization was conducted to validate its structure and to investigate its mechanism of action.Ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was used to confirm the molecular weight,amino acid coverage,and disulfide bond arrangement of the monoclonal antibodies,and its interaction with CD47 was studied by blocking assay.Structural confirmation showed that the measured molecular weight of the monoclonal antibodies was consistent with theoretical molecular weight,The amino acid coverage was 100%and disulfide bond pairing was consistent with the theoretical prediction.Blockade experiments demonstrated that the targeted monoclonal antibody could bind to CD47-his and block the interaction between CD47-his and signal regulatory protein alpha(SIRPα).The molecular structure of the targeted monoclonal antibodies was consistent with its theoretical design structure,which was capable of specifically binding to CD47 and blocking the CD47-SIRPα signaling axis,indicating potential anti-tumor effects.

关 键 词：CD47 结构表征 阻断作用 超高效液相色谱串联质谱 

分 类 号：R392[医药卫生—免疫学]