检测肿瘤McAb新技术DELFIA的建立

The Development of DELFIA with Novel Tumor Monoclonal Antibodies E-11 and G-4

DELFIA(dissociation enhancement lanthanide fluoroimmunoassay)是近年来发展的最新一代免疫分析技术,其全名为解离放大镧系荧光免疫分析法或Tr-FIA(time-resolved fluoroimmuno-assay),即时间分辨荧光免疫分析法,其与传统放射免疫法比较具有更高的灵敏度和精确度.作者从8611卵巢癌细胞株获得McAb F_(11)、C_4、F_(10)、H_1-F_8及H_1-F_7,并分别成功地建立了前4种McAb的DELFIA,本文报告F_(11)及G_4 DELFIA的建立.

Monoclonal antibodies E-11 and G-4 were purified by Sepharose 4B column chromatography and labelled with europium as tracer, The solid-phase antibodies were prepared by attaching pure E-11 and G-4 to polysterene strips.The supoenatant of 8611 cell culture or perchloric acid extracts from ovarian cancer tissues provided standard antigens. The solid-phase sandwich-type DELFIAs for monoclonal antibodies E-11 and G-4 were thus established, and the resulting standard curves were extremely steep. The ratio of CPS in high-concentration tubes to CPS in the zero tube exceeded 300:1, indicating the very high sensitivity and accuracy of this method