靶向人表皮生长因子受体3治疗性单克隆抗体的制备及筛选Preparation and screening of monoclonal antibodies targeting human epidermal growth factor receptor 3
李翱翔;梁红远;祝婧烨;吴丽娜;黄海武;彭波;郭锦林;赵鑫;邱建华;陆瑾;徐帆洪;瞿爱东;李伟民;
摘要(Abstract)：
目的制备并筛选靶向人表皮生长因子受体3(human epidermal growth factor receptor 3,Her3)的单克隆抗体。方法用CHO系统克隆表达Her3的胞外区,获得重组蛋白h GH-Her3-ECD,用该重组蛋白免疫BALB/c小鼠,并经常规细胞融合杂交瘤技术制备单克隆抗体。对制备的单克隆抗体进行抑制Her3与其配体HRG的结合试验、亲和力测定、亚型鉴定、结合抗原结构域的确定、识别肿瘤细胞表面Her3抗原的FACS分析及对A431细胞增殖抑制分析。结果获得100多株靶向Her3的单克隆抗体,其中927、1002、1050、993、1044、1056株具有较强的抑制Her3与HRG结合活性,均为Ig G1型,亲和力较好。927、1002、1050株与Her3第Ⅰ结构域结合,993、1044、1056株与Her3第Ⅲ结构域结合,均能结合A431和Bx Pc3表面的Her3分子,可明显抑制A431细胞的增殖。结论获得的靶向Her3单克隆抗体基本满足开发治疗性单抗的要求,具有成为治疗性单克隆抗体的潜在条件。
关键词(KeyWords)： 人表皮生长因子受体3;单克隆抗体;肿瘤
基金项目(Foundation): 上海市2015年度“科技创新行动计划”生物医药领域科技支持项目(15431905100)
作者(Authors): 李翱翔;梁红远;祝婧烨;吴丽娜;黄海武;彭波;郭锦林;赵鑫;邱建华;陆瑾;徐帆洪;瞿爱东;李伟民;
DOI: 10.13200/j.cnki.cjb.001500
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