生物素-亲和素体系测定雌酮

Abstract

雌酮与牛血清白蛋白共价结合,合成雌酮的完全抗原.利用此抗原免疫小鼠,通过细胞融合技术制备了雌酮的单克隆抗体经纯化表征知,抗本是IgGl型,相对分子量为164000,与固定抗原的亲和常数为8 2×108L/mol. 以生物素化的羊抗鼠免疫球蛋白及辣根过氧化物酶标记的链亲和素为标记体系,通过竞争抑制的方式测定游离的雌酮.结果表明:雌酮在10～10000 ng/mL内呈线性关系

Estrone (E-1) was bound to bovine serum albumin (BSA) to get a complete antigen (E-1 - BSA). A monoclonal antibody (McAb) specific for E-1 was then produced by cell fusion after BALB/c mouse had been immunized with E-1 - BSA. Analysis revealed that anti - E-1 McAb (El - McAb) was of the IgG1 type, and its molecular weight was 164 000 daltons. The affinity constant of E-1 - McAb reacting with coated complete antigen was 8.2 x 10(8) L/mol. Biotin - streptavidin complex was used for the determination of E-1. The calibration curve of free E-1 covered a range of 10 similar to 10 000 pg/mL.