Chu Xia, Fu Xin, Chen Ke, Shen Guo-Li, Yu Ru-Qin
Chemistry and Chemical Engineering College, Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education), Hunan Normal University, Changsha 410081, PR China.
Biosens Bioelectron. 2005 Mar 15;20(9):1805-12. doi: 10.1016/j.bios.2004.07.012.
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 anodic 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 1.66 ng ml(-1) to 27.25 microg ml(-1) in a logarithmic plot. A detection limit as low as 1 ng ml(-1) (i.e., 6 x 10(-12) M) human IgG was achieved, which is competitive with colorimetric enzyme linked immuno-sorbent assay (ELISA) or with immunoassays based on fluorescent europium chelate labels. The high performance of the method is attributed to the sensitive ASV determination of silver (I) at a glassy-carbon electrode (detection limit of 5 x 10(-9) M) and to the catalytic precipitation of a large number of silver on the colloidal gold-labeled antibody.
基于胶体金标记上银的沉淀,开发了一种新型灵敏的电化学免疫分析方法。在酸性溶液中银金属溶解后,通过玻碳电极上的阳极溶出伏安法(ASV)间接测定银。以免疫球蛋白G(IgG)作为模型,对该方法进行了非竞争性异相免疫分析评估。研究并优化了相关实验变量,包括抗原与抗体的反应时间、胶体金标记抗体的稀释比例以及阳极溶出操作参数对峰电流的影响。在对数图中,阳极溶出峰电流在1.66 ng ml(-1)至27.25 μg ml(-1)范围内与IgG浓度呈线性关系。实现了低至1 ng ml(-1)(即6×10(-12) M)的人IgG检测限,这与比色酶联免疫吸附测定(ELISA)或基于荧光铕螯合物标记的免疫分析方法具有竞争力。该方法的高性能归因于在玻碳电极上对银(I)的灵敏ASV测定(检测限为5×10(-9) M)以及在胶体金标记抗体上大量银的催化沉淀。