Title基于温度敏感水凝胶的雌二醇荧光免疫分析
Other TitlesFluoroimmunoassay of estradiol based on the thermo-sensitive hydrogel
Authors王永成
张新祥
李元宗
常文保
Affiliation北京大学化学与分子工程学院
北京大学化学与分子工程学院 北京100871
北京100871
Keywords雌二醇
聚异丙基丙烯酰胺
荧光免疫分析
estradiol
poly-N-isopropylacrylamide
fluoroimmunoassay
Issue Date2002
Publisher高等学校化学学报
Citation高等学校化学学报.2002,(05),792-795.
Abstract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.
URIhttp://hdl.handle.net/20.500.11897/78283
ISSN0251-0790
DOI10.3321/j.issn:0251-0790.2002.05.007
IndexedSCI(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)
Appears in Collections:化学与分子工程学院

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