高等学校化学学报

高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (5): 792.

• 研究简报 • 上一篇    下一篇

基于温度敏感水凝胶的雌二醇荧光免疫分析

王永成, 张新祥, 李元宗, 常文保   

  1. 北京大学化学与分子工程学院, 北京 100871
  • 收稿日期:2001-05-21 出版日期:2002-05-24 发布日期:2002-05-24
  • 通讯作者: 常文保(1939年出生),男,教授,博士生导师,从事生化免疫分析研究.E-mail:dxhx@chem.pku.edu.cn E-mail:dxhx@chem.pku.edu.cn
  • 基金资助:

    国家自然科学基金(批准号:20075001)资助.

Fluoroimmunoassay of Estradiol Based on the Thermo-sensitive Hydrogel

WANG Yong-Cheng, ZHANG Xin-Xiang, LI Yuan-Zong, CHANG Wen-Bao   

  1. College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China
  • Received:2001-05-21 Online:2002-05-24 Published:2002-05-24

PDF (PC)

被引次数

17

摘要： 雌二醇完全抗原(Ag)与异丙基丙烯酰胺(NIPA)共聚,可得水凝胶－抗原结合物(pNIPA－Ag),pNI－PA-Ag与游离的雌二醇(E2)竞争有限的荧光标记的单抗McAb-F,利用水凝胶温度相变的性质,分离高分子免疫复合物,并测定其荧光信号.结果表明,游离E2在10～625ng/mL范围内呈线性关系,样品的回收率为88.9%～102.0%.

关键词: 雌二醇, 聚异丙基丙烯酰胺, 荧光免疫分析

Abstract: An indirect competitive fluoroimmunoassay of estradiol was developed based on the thermo-sen-sitive hydrogel, poly-JV-isopropylacrylamide(pNIPA).Antigen was conjugated to pNIPAto form antigen bound thermally reversible hydrogel (pNIPA-Ag) after the antigen was synthesized by covalent bonding estradiol to bovine serum albumin.The anti-estradiol monoclonal antibody (McAb) was prepared by an in vivo method with McAb secreting hybridomas.Fluorescein isothiocyanate(FITC) labeled McAb(McAb-F) was prepared by bonding FITCto purified McAb.Then free estradiol reacted with limited amount of McAb-Fcompeting with pNIPA-Ag.Separation of the immune complex, pNIPA-Ag-McAb-F, was carried out by centrifuging and precipitating at a temperature above critical solution temperature of the hydro-gel.After the above immune complex was dissolved, the fluorescence intensity was measured by a Hitachi M-850 fluorospectrometer.The calibration curve of free estradiol covered a range of 10-625 ng/mL, and the detection limit of the presented method for free estradiol was 3.2 ng/mL.The cross-reactions and the recoveries of the immunoassay were also investigated.

Key words: Estradiol, Poly-N-isopropylacrylamide, Fluoroimmunoassay

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