胶体金免疫层析法参数的优化

研究胶体金免疫层析法中几种重要影响因素的参数优化，并以甲基对氧磷为对象进行实验。首先对现有的抗体纯化方法进行优化选择，根据SDS-聚丙烯酰胺凝胶电泳结果确定最佳抗体纯化方法，并用该法对目标抗体甲基对氧磷进行纯化。将纯化后的多克隆抗体与30nm的胶体金溶液进行标记，并对金标记抗体进行纯化。对反应溶液系统进行筛选，确定最合适的重悬液、抗原稀释液以及样品垫处理液。最终对试纸条的灵敏度，样品基质对试纸条的影响进行评测，对于目标检测物的裸眼检测限达到5μg/ml，样品的基质效应对试纸条的性能没有影响。

Optimization of Parameters of Colloidal Golden Immune Chromatography

Methyl paraoxon was subjected to explore the optimal parameters of colloidal gold immune chromatography. First, currently available antibody purification methods were screened to decide the optimal one based on SDS-PAGE. The obtained optimal method was used to purify methyl paraoxon antibodies. Then, the labeling was completed through the reaction between purified polycolonial antibodies and colloidal gold solutions with 30 nm of particle size. Reaction solutions were further screened to determine the optimal re-suspension, dilution of antigens and treatment solutions for sample pads. Finally, the dipsticks with colloidal gold were developed, which exhibited simple and rapid detection of methyl paraoxon. The final performance on these test strips was evaluated and the detection limit for targets was 5μg/ml using naked eye.