抗癌胚抗原噬菌体单链抗体库的构建和筛选鉴定  

Construction and screening of a phage single-chain fragment variable library against carcinoembryonic antigen

作　　者：陈团结 梁红远 陈湛昊 王明辉 瞿爱东 陈哲文 Chen Tuanjie;Liang Hongyuan;Chen Zhanhao;Wang Minghui;Qu Aidong;Chen Zhewen(No.4 Research Laboratory,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 200051,China;No.1 Research Laboratory,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 200051,China;Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 201403,China)

机构地区：[1]上海生物制品研究所有限责任公司第四研究室,上海200051 [2]上海生物制品研究所有限责任公司第一研究室,上海200051 [3]上海生物制品研究所有限责任公司,上海201403

出　　处：《国际生物制品学杂志》2024年第4期191-196,共6页International Journal of Biologicals

摘　　要：目的以癌胚抗原相关黏附分子5(carcinoembryonic antigen-related cell adhesion molecule 5,CEA)作为靶点,构建靶向于CEA的噬菌体单链抗体(single-chain fragment variable,ScFv)免疫库,并通过筛选获得靶向该抗原的抗体。方法通过分子克隆的方法将小鼠脾脏中的抗体序列与噬菌体质粒构建获得重组噬菌体质粒,并经电转化导入大肠埃希菌TG1菌株中,再使用M13KO7辅助噬菌体拯救的方法,从TG1菌株中获得多样化的噬菌体抗体库。使用体外噬菌体库固相亲和筛选的方法,以CEA蛋白为靶点筛选获得高亲和力抗体。结果成功构建了鼠源抗CEA噬菌体ScFv免疫库,经计算库容量为1.02×10^(8) CFU,并通过筛选获得了靶向CEA的ScFv。结论利用噬菌体展示技术,筛选了靶向CEA的抗体,为该靶点抗体的筛选奠定基础。Objective To construct a single-chain fragment variable(ScFv)immune library targeting carcinoembryonic antigen-related cell adhesion molecule 5(CEA),and to obtain antibodies targeting CEA by screening..Methods The recombinant phage plasmids were constructed by cloning the antibody sequences from mouse spleen into phage plasmids.The plasmids were then electrically transformed into E.coli strain TG1,and rescued by M13KO7 helper phage to obtain a variety of phage antibody libraries from TG1 strain.Using in vitro phage library solid-phase affinity screening method and CEA protein as target,high-affinity antibodies were screened.Results A murine-derived anti-CEA phage ScFv immune library was successfully constructed,and the library capacity was calculated to be 1.02×10^(8) CFU.ScFv targeting CEA was obtained by screening.Conclusion The anti-CEA antibody is screened by the phage display technique,laying foundation for the screening of anti-CEA antibody.

关 键 词：噬菌体库 单链抗体 癌胚抗原 肿瘤 

分 类 号：R-5[医药卫生]