Poly(o-phenylenediamine)-carried nanogold particles as signal tags for sensitive electrochemical immunoassay of prolactin

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• 作者：Huafeng Chen^a ; ^b ; Yuling Cui^a ; ^b ; Bing Zhang^a ; ^b ; Bingqian Liu^a ; ^b ; Guonan Chen^a ; ^b ; Dianping Tang^a ; ^b ; ^{dianping.tang@fzu.edu.cn}
• 关键词：Electrochemical immunoassay ; Nanogold-assembled poly(o-phenylenediamine) microspheres ; Nanolabels ; Prolactin
• 刊名：Analytica Chimica Acta
• 出版年：2012
• 期刊代码：2_00032670
• 类别：ch
• 出版时间：30 May, 2012
• 卷：728
• 期：Complete
• 页码：18-25
• 文件大小：1503 K

摘要

A novel class of redox-active molecular tags, poly(o-phenylenediamine)-carried nanogold particles (GPPDs), was first synthesized and functionalized with horseradish peroxidase-anti-prolactin conjugates (HRP-anti-PRL). Thereafter, a specific sandwich-type electrochemical immunoassay was designed for determination of prolactin (PRL) by using GPPD-labeled HRP-anti-PRL conjugates as molecular tags on anti-PRL antibody-modified glassy carbon electrode. Compared with pure gold nanoparticles and poly(o-phenylenediamine) microspheres, the as-prepared GPPDs increased the surface coverage of the nanostructures, and enhanced the immobilization amount of biomolecules. Several labeling protocols compromising GPPD-labeled HRP-anti-PRL, nanogold particles-labeled HRP-anti-PRL and poly(o-phenylenediamine) microspheres-labeled HRP-anti-PRL, were investigated for detection of PRL, and improved analytical features were obtained with the GPPD-based strategy. With the GPPD labeling method, dependence of the electrochemical signals on the incubation time and pH of the assay solution were also studied. The strong attachment of HRP-anti-PRL to the GPPDs resulted in a good repeatability and intermediate reproducibility down to 9.8%. The dynamic concentration range spanned from 0.5 to 180 ng mL^鈭? PRL with a detection limit of 0.1 ng mL^鈭? at the 3S_blank level. No significant differences at the 95%confidence level were encountered in the analysis of 10 spiked blank cattle serum samples between the developed immunoassay and enzyme-linked immunosorbent assay method for determination of PRL.