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	基于温度敏感水凝胶的雌二醇荧光免疫分析; Fluoroimmunoassay of estradiol based on the thermo-sensitive hydrogel
	王永成 ; 张新祥 ; 李元宗 ; 常文保
	2002
关键词	雌二醇 聚异丙基丙烯酰胺 荧光免疫分析 estradiol poly-N-isopropylacrylamide fluoroimmunoassay
英文摘要	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％．; An indirect competitive fluoroimmunoassay of estradiol was developed based on the thermo-sensitive hydrogel, poly-N-isopropylacrylamide(pNIPA). Antigen was conjugated,to pNIPA to 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 FITC to purified McAb. Then free estradiol reacted with limited amount of McAb-F competing 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 hydrogel. 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.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000175831000010&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; SCI(E); EI; 中文核心期刊要目总览(PKU); 中国科技核心期刊(ISTIC); 中国科学引文数据库(CSCD); 6; 05; 792-795; 23
语种	中文
出处	万方 ; 知网 ; SCI
出版者	高等学校化学学报
内容类型	其他
源URL	[http://hdl.handle.net/20.500.11897/78283]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	王永成,张新祥,李元宗,等. 基于温度敏感水凝胶的雌二醇荧光免疫分析, Fluoroimmunoassay of estradiol based on the thermo-sensitive hydrogel. 2002-01-01.

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