Use of <scp>l</scp>-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens
作者:George I. Hatzidakis、Aristidis M. Tsatsakis、Elias K. Krambovitis、Apostolos Spyros、Sergei A. Eremin
DOI:10.1021/ac011051x
日期:2002.6.1
the hapten attached to the alpha-aminogroup of L-lysine and fluorescein to the e-amino group exhibited at least a 5-fold increased sensitivity when compared to the previously reported ethylenediamine-based tracer (2,4D-EDA-F). The isomeric structure (hapten attached to the e-amino and fluorescein to the alpha-amino group) appeared 7.6 times less sensitive, and all other alternative structures exhibited
荧光偏振免疫测定法(FPIA)是一种方便的均相测定法,由于其灵敏度低和基质效应低,在环境分析中的使用受到限制。我们选择了除草剂2,4D和2,4,5T来使用固相化学合成新的基于L-赖氨酸的荧光示踪剂。另外,制备了2、4、5T的三种不同的免疫原用于免疫和抗体产生。新的示踪剂和抗体适用于FPIA。与先前报道的基于乙二胺的示踪剂(2,4D-EDA-F)相比,半抗原连接到L-赖氨酸的α-氨基和荧光素连接到e-氨基的示踪剂显示出至少5倍的灵敏度提高。 。异构体结构(半抗原连接到e-氨基,荧光素连接到α-氨基)灵敏度降低了7.6倍,其他所有替代结构的灵敏度都更低。针对单克隆抗2,4D抗体E2 / G2和多克隆抗2,4,5T抗体证实了该观察结果。与新型示踪剂相比,2,4D-EDA-F与E2 / G2的亲和常数高8.1倍,这表明基于L-赖氨酸的示踪剂具有更特殊的性质,使用该示踪剂会导致更灵敏的测定。这种类型