吗啡的控温相分离胶体金标记免疫分析

Abstract

首次将胶体金作为标记物引入相变免疫分析中 ,建立了吗啡免疫分析的新方法。该分析方法是将吗啡抗体与温度敏感型水凝胶偶联 ,胶体金标记吗啡全抗原 ,再进行相变分离 ,最后用可见吸收光谱仪进行检测。本方法的两个重要特点是 :( 1)利用温敏水凝胶的相变特性进行均相反应和异相分离 ,缩短了免疫反应时间 ,简化了步骤 ;( 2 )利用胶体金快速稳定吸附蛋白质的特性进行标记 ,利用胶体金本身的颜色 ,可进行定性检测 ,或者用常规的可见吸收光谱仪进行定量检测 ,对仪器要求简单 ,适用于临床检测的要求。

A novel immunoassay method for the determination of morphine has been developed by introducing colloid gold to this phase separation system. The colloid gold as the marker is combined with the morphine antigen. The morphine antibody is conjugated to poly-N-isopropylacrylamide(PNIP, a kind of hydrogel) forming thermally reversible PNIP-Ab(MP). After competitive reaction of colloid gold combined morphine and free morphine with limited PNIP-Ab(MP), the content of free morphine in sample can be quantified indirectly through the quantity of colloid gold, which absorbs the 530 nm-wavelength light. This method has two main characters: 1. A phase transformation of PNIP will take place when its temperature increases or decreases through 31 degreesC, thus home-phase action and hetero-phase separation can be possible. 2. The colloid gold has the character of absorbing protein quickly and firmly, so it can be used as the tracer in immune reaction either combined with antigen or combined with antibody. The colloid gold can. be quantified with an inexpensive instrument in a short time, so it accords with the needs of routine measurement.