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基于酶促生物催化沉淀的纳米金功能化 g-C3N4 纳米杂化材料用于灵敏的前列腺特异性抗原的电化学免疫分析

Nanogold-functionalized g-C3N4 nanohybrids for sensitive impedimetric immunoassay of prostate-specific antigen using enzymatic biocatalytic precipitation.

机构信息

Department of Urology, Jinling Hospital, Medical School of Nanjing University, Nanjing University, Nanjing, Jiangsu Province 210002, PR China.

Department of Urology, Jinling Hospital, Medical School of Nanjing University, Nanjing University, Nanjing, Jiangsu Province 210002, PR China.

出版信息

Biosens Bioelectron. 2016 Nov 15;85:212-219. doi: 10.1016/j.bios.2016.04.102. Epub 2016 May 6.

DOI:10.1016/j.bios.2016.04.102
PMID:27179136
Abstract

This work reports on a new impedimetric immunosensing strategy for sensitive detection of prostate-specific antigen (PSA) in biological fluids. The assay was carried out on monoclonal anti-PSA capture antibody-modified glassy carbon electrode with a sandwich-type detection format. Gold nanoparticles-decorated g-C3N4 nanosheets (AuNP/g-C3N4), synthesized by the wet-chemistry method, were utilized for the labeling of polyclonal anti-PSA detection antibody and horseradish peroxidase (HRP). Upon target PSA introduction, the sandwiched immunocomplex could be formed between capture antibody and detection antibody. Followed by the AuNP/g-C3N4, the labeled HRP could catalyze 4-choloro-1-naphthol into benzo-4-chlorohexadienone. The as-generated insoluble product was coated on the electrode surface, thus increasing the Faradaic impedance of Fe(CN)6(4-/3)(-) indicator between the solution and the base electrode. Under the optimal conditions, the impedance increased with the increasing target PSA in the sample, and exhibited a wide linear range from 10pgmL(-1) and 30ngmL(-1) with a detection limit of 5.2pgmL(-1). A repeatability and intermediate precision of <14% was accomplished. The specificity and method accuracy in comparison with commercial PSA ELISA kit for analysis of human serum specimens were relatively satisfactory.

摘要

本工作报道了一种新的基于阻抗的免疫传感策略,用于生物流体中前列腺特异性抗原(PSA)的灵敏检测。该测定采用单克隆抗 PSA 捕获抗体修饰的玻碳电极,采用三明治型检测模式进行。通过湿化学法合成的金纳米粒子修饰的 g-C3N4 纳米片(AuNP/g-C3N4)用于多克隆抗 PSA 检测抗体和辣根过氧化物酶(HRP)的标记。引入靶 PSA 后,可在捕获抗体和检测抗体之间形成夹心免疫复合物。随后加入 AuNP/g-C3N4,标记的 HRP 可将 4-氯-1-萘酚催化生成苯并-4-氯己二烯酮。生成的不溶性产物覆盖在电极表面,从而增加了溶液和基底电极之间的 Fe(CN)6(4-/3)(-)指示剂的法拉第阻抗。在最佳条件下,阻抗随样品中目标 PSA 的增加而增加,线性范围从 10pgmL(-1)到 30ngmL(-1),检测限为 5.2pgmL(-1)。重复性和中间精密度小于 14%。与用于分析人血清标本的商业 PSA ELISA 试剂盒相比,该方法具有较好的特异性和方法准确性。

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