烯二炔抗癌抗生素单克隆抗体的制备及初步应用  

A MONOCLONAL ANTIBODY DIRECTED AGAINST AN ENEDIYNE ANTITUMOR ANTIBIOTIC AND ITS PRELIMINARY APPLICATION

作　　者：周春水[1,2] 徐琳娜[1,2] 江敏 甄永苏[1,2] 

机构地区：[1]中国医学科学院 [2]中国协和医科大学医药生物技术研究所

出　　处：《药学学报》1997年第1期28-32,共5页Acta Pharmaceutica Sinica

基　　金：国家"863"计划资助

摘　　要：抗癌抗生素Ｃ１０２７分子由一个含１１０个氨基酸的蛋白和一个含烯二炔结构的发色团非共价结合而成。发色团是Ｃ１０２７的活性部分，易被紫外线照射灭活，蛋白则对发色团有保护作用。以紫外灭活的Ｃ１０２７为抗原，用杂交瘤技术，制备了小鼠抗Ｃ１０２７单克隆抗体Ｆ９。免疫印迹证实单抗Ｆ９能特异性识别Ｃ１０２７，该单抗对Ｃ１０２７与紫外线灭活Ｃ１０２７的亲和力无显著差别。单抗Ｆ９对Ｃ１０２７的亲和常数为２２±０４７×１０７Ｌ·ｍｏｌ－１，其亚型为ＩｇＧ１。克隆形成测定结果表明，单抗Ｆ９对Ｃ１０２７的细胞毒性无明显影响，提示该单抗没有对Ｃ１０２７的发色团产生封闭效应。ＥＬＩＳＡ与Ｗｅｓｔｅｒｎｂｌｏｔ测试说明单抗Ｆ９能与裁短的Ｃ１０２７基因工程肽发生特异性结合。提示单抗Ｆ９可用于Ｃ１０２７基因工程的研究，并可能用于研究Ｃ１０２７的代谢与分布及筛选大分子抗肿瘤抗生素等方面。C1027, composed of an enediyne chromophore and an apoprotein of 110 amino acid residues, is a new highly potent macromolecular antitumor antibiotic. In order to prepare monoclonal antibodies (McAb) against C1027 by hybridoma technique, natural C1027 was inactivated by ultraviolet and coupled with human serum albumin, then immunized BALB/c mice. After cell fusion and screening by ELISA, a hybridoma secreting anti C1027 McAb designated as F9 was obtained. McAb F9 specifically reacted with C1027 as determined by immunoblot assay. No obvious difference was observed between the reactivity of McAb F9 to natural C1027 and that of McAb F9 to ultraviolet inactivated C1027. This result indicates that the ultraviolet sensitive chromophore of C1027 may not participate in the epitope for McAb F9. The isotype of F9 is IgG 1 and its affinity constant was found to be (2 2±0 47)×10 7 L·mol -1 according to Beatty′s method. By clonogenic assay, McAb F9 neither affected the cytotoxicity of C1027 to KB cells nor blocked the activity of the chromophore. McAb F9 also specifically reacted with the recombinant truncated C1027 apoprotein in which 16 amino acid residues at the C terminus were deleted. This study suggests that F9 is a valuable McAb for the research of C1027 apoprotein engineering, C1027 related immunoconjugates and screening of new macromolecular antitumor substances with homology to the protein part of C1027.

关 键 词：抗癌抗生素 C1027 单克隆抗体 烯二炔 制备 

分 类 号：R392－33[医药卫生—免疫学]