时间分辨荧光免疫分析方法的光谱研究

时间分辨荧光免疫分析法是用三价稀土离子及其螯合剂作为示踪物 ,标记蛋白质、激素、抗体、核酸探针或生物活性细胞 ,待反应体系 (如 :抗原抗体免疫反应、生物素亲合反应、核酸探针杂交反应、靶细胞与效应细胞的杀伤反应等 )发生后 ,用时间分辨荧光技术测定反应体系中分析物的浓度 ,达到定量分析的目的。它之所以能够继放射性同位素标记、酶标记、化学发光、电化学发光后成为一种更新、更灵敏的检测方法 ,主要取决于它所用标记物三价稀土离子螯合物独一无二的物理及化学性质。主要报导了对使用的长寿命荧光团Eu3 螯合物的光谱研究结果 ,时间分辨技术及荧光增强技术的原理。实验表明 :选择 336～ 337nm的激发波长 ,有利于Eu3 的配位二酮体的激发及能量转移。

Spectroscopic Evaluation of Time-Resolved Fluoroimmunoassay

The lanthanide trivalence ion and its chelates are used for marking substance in Time-Resolved Fluorescence Immunoassay (TRFIA), marking protein, hormone, antibody, nucleic acid probe or biologic alive cell, to measure the concentration of the analysis substance inside the reaction system with time-resolved fluorometry after the reaction system occurred, and attain the quantitative analysis's purpose. TRFIA has therefore become a kind of new and more sensitive measurement method after radioisotope marking, enzymatic marking, chemiluminescence, electrochemiluminescence, primarily depending on the special physics and chemistry characteristics of lanthanide trivalence ion and its chelates. In this paper, the result of spectroscopic evaluation of europium trivalence ion and its chelate, and the principle of time-resolved technology and fluorescence-enhanced technology are reported. At the same time, the experiment shows that the excitation wavelength chosen between 336 and 337 nm benefits the excitation and the energy transfer of chelate diketone of europium trivalence ion.