基于银染色放大和免疫胶体金的金属免疫分析  

Metalloimmunoassay based on silver-staining enhancement and immuno-colloidal gold

作　　者：傅昕[1] 向知峰[1] 楚霞[1,2] 

机构地区：[1]湖南师范大学化学化工学院,湖南长沙410081 [2]化学与生物传感及化学计量学国家重点实验室,湖南长沙410082

出　　处：《化学传感器》2004年第3期39-42,共4页Chemical Sensors

基　　金：国家自然科学基金(Nos:20105007)资助项目

摘　　要：提出了一种新的、高灵敏的基于银染色放大和免疫胶体金技术的金属免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过纳米金对银离子的催化还原在金标抗体上沉积大量的金属银,用酸溶解后,在玻碳电极上用阳极溶出伏安法(ASV)对银离子进行检测,溶出峰电流的大小间接与待分析物人免疫球蛋白G(IgG)的浓度成正比。对阳极溶出分析的测定参数如沉积电位、富集时间和免疫分析的一些条件如抗原-抗体反应时间、金标抗体稀释度等进行了优化。阳极溶出峰电流与IgG浓度在1.66μg/L～27.25 mg/L范围内呈良好的线性关系,检测下限为1.1μg/L。A novel, sensitive electrochemical immunoassay has been developed based on the precipitation of silver on colloidal gold labels which, after silver metal dissolution in an acidic solution, was indirectly determined by anodic stripping voltammetry (ASV) at a glassy-carbon electrode. The method was evaluated for a noncompetitive heterogeneous immunoassay of an immunoglobulin G (IgG) as a model. The influence of relevant experimental variables, including the reaction time of antigen with antibody, the dilution ratio of the colloidal gold-labeled antibody and the parameters of the anodie stripping operation, upon the peak current was examined and optimized. The anodic stripping peak current depended linearly on the IgG concentration over the range of 1.66μg/L to 27.25 mg/L and a detection limit as low as 1.1μg/L was achieved.

关 键 词：银染色 免疫分析方法 抗体 人免疫球蛋白 免疫胶体金 测定参数 免疫反应 银离子 阳极 放大 

分 类 号：Q503[生物学—生物化学]