转化生长因子-β_1单克隆抗体TB21的生物学特性(英文)  

BIOLOGICAL CHARACTERIZATION OF A MONOCLONAL ANTIBODY TB21 AGAINST HUMAN TRANSFORMING GROWTH FACTOR-BETA 1

作　　者：施渭康[1] 叶庆炜 包林平[1] 

机构地区：[1]中国科学院上海细胞生物学研究所 [2]YES生物技术公司

出　　处：《实验生物学报》1993年第2期141-150,共10页Acta Biologiae Experimentalis Sinica

摘　　要：转化生长因子-β_1(TGF-β_1)是转化生长因子家族中的一个重要成员,具有多种生物学功能,特别是调节细胞的生长和分化,参与细胞外基质合成,创伤愈合和胚胎形态发生等生物学过程。近年来,发展了一些抗TGF-β_1多克隆抗体和个别重组人TGF-β_1的单克隆抗体,对研究各种细胞的TGF-β_1的合成,定位和其它生物学效应发挥了作用。本文报道了抗人血小板TGF-β_1单克隆抗体TB 21的制备及其主要生物学特性,尤其是它对TGF-β_1免疫识别的专一性,以及调变对TGF-β_1敏感细胞的生长和增殖抑制的活性。结果表明,TB_(21)单克隆抗体为IgG_1亚型;对TGF-β_1有较高亲和力,亲和常数(Kaff)为1.47×10~8M^(-1);Westernblot 显示TB_(21)抗体能专一地结合25 Kd 成熟型分子和其12.5 Kd 的单体分子,对CCL/64细胞生长抑制和对NRK-49 F 成纤维细胞软琼脂集落形成的鉴定都证明TB21单抗对TGF-β_1的生物学效应有专一性的中和作用,可用于TGF-β_1的有关生物学研究。A hybridoma TB21 clone was derivedfrom fusion between Sp2/0-Ag 14 myelo-ma cells and spleen cells of BALB/c miceimmunized with transforming growth fac-tor-β_1(TGF-β_1)purified from humanplatelets.The TB21 clone was identifiedto produce monoclonal antibody with IgGlsubclass and had sufficient titer for immu-noreactivity to both human platelets-deri-ved TGF-β_1 and recombinant human TGF-β_1 by enzemy-linked immunosorbentassay(ELISA).Western blotting studiesdemonstrated that two immunoreactive ba-nds corresponding to 25 Kda and 12.5 Kdamoleculxs were observed in the sample ofacid/ethanol extracts from human platelets.The affinity constant(Kaff)was determi-ned as 1.47×10~8 M^(-1) with non-competi-tive ELISA.Moreover,using bioassay forthe effects of TGF-β_1 on the growth ofmink lung epithelial cells(CCL/64 cellline)and fibroblast cells(NRK 49F cellline),TB21 IgG was shown to be able toneutralize the action of TGF-β_1 on thegrowth of these target cells.Therefore,this monoclonal antibody may be a usefulprobe for studying the growth modulatoryactivity of TGF-β_1 in a variety of cellsand tissues.

关 键 词：单克隆抗体 转化生长因子 

分 类 号：Q28[生物学—细胞生物学]