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化学学报
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化学学报 ›› 2001, Vol. 59 ›› Issue (10): 1609-1613. 上一篇    下一篇

研究论文

生物素-亲和素体系测定雌酮

王永成;李元宗;常文保;郭振泉;慈云祥   

  1. 北京大学化学与分子工程学院;北京大学生命科学中心
  • 发布日期:2001-10-15

Determination of esterone using biotin-streptavidin complex

Wang Yongcheng;Li Yuanzong;Chang Wenbao;Guo Zhenquan;Ci Yunxiang   

  • Published:2001-10-15

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被引次数

9 | 2

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，抗体是IgG1型，相对分子量为164000,与固定抗的亲和常数为8.2×10^8L/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10～10000pg/mL内呈线性关系。

关键词: 生物素, 雌酮, 单克隆抗体, 免疫测定

Estrone (E1) was bound to bovine serum albumin (BSA) to get a complete antigen (E1-BSA). A monoclonal antibody (McAb) specific for E1 was then produced by cell fusion after BALB/c mouse had been immunized with E1-BSA. Analysis revealed that anti-E1 McAb(E1-McAb) was of the IgG1 type, and its molecular weight was 164 000 datons. The affinity constant of E1-McAb reacting with coated complete antigen was 8.2×10^8L/mol. Biotin-streptavidin complex was used for the determination of E1. The calibration curve of free E1 covered a range of 10～10 000 pg/mL.

Key words: BIOTIN, FOLLICULIM (=ESTRONE), MONOCLONAL ANTIBODY, IMMUNOASSAY

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