提出了一种基于纳米金标记的阳极溶出伏安免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过物理吸附将兔抗人免疫球蛋白 A(IgA)抗体固定于微孔板上,与相应抗原 IgA 发生免疫反应后,再通过夹心模式捕获相应的纳米金标...提出了一种基于纳米金标记的阳极溶出伏安免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过物理吸附将兔抗人免疫球蛋白 A(IgA)抗体固定于微孔板上,与相应抗原 IgA 发生免疫反应后,再通过夹心模式捕获相应的纳米金标记的羊抗人 IgA 抗体,然后再与金标羊抗人 IgA 抗体和金标兔抗羊二抗形成的免疫复合物反应,在微孔板上引入大量的纳米金,将金溶解后,在碳糊电极上用阳极溶出伏安法(ASV)对金离子进行检测,溶出峰电流的大小间接与待分析物 IgA 的浓度成正比。对免疫分析的一些实验条件进行了优化。阳极溶出峰电流与 IgA 的浓度在0.018 mg/L～181 mg/L 范围内呈良好的线性关系,检测下限为10μg/L。将该分析方法应用于人血清中 IgA 浓度的测定,取得了满意结果。展开更多
Based on MEMS technology,immunosensor with an'Au,Pt,Pt'three-microelectrode system enclosed in a SU-8 micro pool was fabricated.Employing SAMs technique,the Au electrode was modified by cysteamine(Cys)to assem...Based on MEMS technology,immunosensor with an'Au,Pt,Pt'three-microelectrode system enclosed in a SU-8 micro pool was fabricated.Employing SAMs technique,the Au electrode was modified by cysteamine(Cys)to assemble gold nanopanicles(nanogold)layer,subsequently,a layer of protein G(PG)was immobilized on nanogold layer to further capture antibody orientedly.Compared with the immunosensors using bulky gold electrode and direct PG binding to electrode immobilization technique for antibody,it has attractive advantages,such as miniaturization,good compatibility,broad linear range for human immunoglobulin(HIgG)and easy to be designed into array.展开更多
The interaction between gold nanoparticles and proteins such as bovine serum albumin and immunoglobulin G under the condition of different pH values was studied based on the measurement of zeta potential and fluoresce...The interaction between gold nanoparticles and proteins such as bovine serum albumin and immunoglobulin G under the condition of different pH values was studied based on the measurement of zeta potential and fluorescence quenching of the proteins before and after proteins were bound with gold nanoparticles. Aggregations were found in gold colloid aqueous solution after addition of proteins by TEM characterization and UV-Vis spectroscopy deterruination. The results showed that the values of zeta potential were quite different, the binding constant Kb and stoichiometry n were slightly increased with the increase of pH value. In conclusion, two factors could affect markedly the interaction between gold nanoparticles and proteins, that is, surface charge and the coordination effect between gold nanoparticles and indole group of the tryptophan residue of proteins.展开更多
文摘提出了一种基于纳米金标记的阳极溶出伏安免疫分析方法。免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过物理吸附将兔抗人免疫球蛋白 A(IgA)抗体固定于微孔板上,与相应抗原 IgA 发生免疫反应后,再通过夹心模式捕获相应的纳米金标记的羊抗人 IgA 抗体,然后再与金标羊抗人 IgA 抗体和金标兔抗羊二抗形成的免疫复合物反应,在微孔板上引入大量的纳米金,将金溶解后,在碳糊电极上用阳极溶出伏安法(ASV)对金离子进行检测,溶出峰电流的大小间接与待分析物 IgA 的浓度成正比。对免疫分析的一些实验条件进行了优化。阳极溶出峰电流与 IgA 的浓度在0.018 mg/L～181 mg/L 范围内呈良好的线性关系,检测下限为10μg/L。将该分析方法应用于人血清中 IgA 浓度的测定,取得了满意结果。
基金This work is supported by the National Natural Science Foundation of China （Grant No. 90307014）.
文摘Based on MEMS technology,immunosensor with an'Au,Pt,Pt'three-microelectrode system enclosed in a SU-8 micro pool was fabricated.Employing SAMs technique,the Au electrode was modified by cysteamine(Cys)to assemble gold nanopanicles(nanogold)layer,subsequently,a layer of protein G(PG)was immobilized on nanogold layer to further capture antibody orientedly.Compared with the immunosensors using bulky gold electrode and direct PG binding to electrode immobilization technique for antibody,it has attractive advantages,such as miniaturization,good compatibility,broad linear range for human immunoglobulin(HIgG)and easy to be designed into array.
基金Project supported by the National Natural Science Foundation of China （No. 30471599） and Development Foundation of Shanghai Science and Technology （No.03ZR14057）.
文摘The interaction between gold nanoparticles and proteins such as bovine serum albumin and immunoglobulin G under the condition of different pH values was studied based on the measurement of zeta potential and fluorescence quenching of the proteins before and after proteins were bound with gold nanoparticles. Aggregations were found in gold colloid aqueous solution after addition of proteins by TEM characterization and UV-Vis spectroscopy deterruination. The results showed that the values of zeta potential were quite different, the binding constant Kb and stoichiometry n were slightly increased with the increase of pH value. In conclusion, two factors could affect markedly the interaction between gold nanoparticles and proteins, that is, surface charge and the coordination effect between gold nanoparticles and indole group of the tryptophan residue of proteins.
