A novel electrochemical immunosensor for tumor biomarker detection based on three-dimensional, magnetic and electroactive nanoprobes was developed in this study. To fabricate the nanoprobes, negatively charged Fe(3)O(4) nanoparticles (Fe(3)O(4) NPs) and gold nanoparticles (Au NPs) were first loaded on the surface of multiple wall carbon nanotubes (MCNTs) which were functioned with redox-active hemin and cationic polyelectrolyte poly(dimethyldiallylammonium chloride) (PDDA). Using alpha fetoprotein (AFP) as a model analyte, AFP antibody (anti-AFP) was absorbed on the surface of Au NPs, bovine serum albumin (BSA) was then used to block sites against non-specific binding, and finally formed anti-AFP/Au NPs/Fe(3)O(4)/hemin/MCNTs named anti-AFP nanoprobes. When the target antigen AFP was present, it interacted with anti-AFP and formed an antigen-antibody complex on the nanoprobe interface. This resulted in a decreased electrochemical signal of hemin for quantitative determination of AFP when immobilized onto the screen-printed working electrode (SPCE). The results showed that the nanoprobe-based electrochemical immunosensor was sensitive to AFP detection at a concentration of 0.1 to 200 ng·mL(-1) with a detection limit of 0.04 ng·mL(-1), it also demonstrated good selectivity against other interferential substances. The electroactive nanoprobes can be massively prepared, easily immobilized on the SPCE for target detection and rapidly renewed with a magnet. The proposed immunosensor is fast, simple, sensitive, stable, magnet-controlled, nontoxic, label-free and reproducible.

译文

本研究开发了一种基于三维,磁性和电活性纳米探针的用于肿瘤生物标志物检测的新型电化学免疫传感器。为了制造纳米探针,首先将带负电的Fe(3)O(4) 纳米颗粒 (Fe(3)O(4) NPs) 和金纳米颗粒 (Au NPs) 负载在具有氧化还原活性的血红素和阳离子聚电解质聚 (二甲基二烯丙基氯化铵) (PDDA)。以甲胎蛋白 (AFP) 为模型分析物,AFP抗体 (anti-AFP) 被吸收在Au NPs表面,牛血清白蛋白 (BSA) 被用于阻断位点对抗非特异性结合,最后形成了名为抗AFP/Au NPs/Fe(3)O(4)/hemin/MCNTs的抗AFP纳米探针。当目标抗原AFP存在时,它与抗AFP相互作用,并在纳米探针界面上形成抗原-抗体复合物。当固定在丝网印刷的工作电极 (SPCE) 上时,这导致用于定量测定AFP的血红素的电化学信号降低。结果表明,基于纳米探针的电化学免疫传感器在0.1 ~ 200 ng·mL(-1) 浓度下对AFP检测敏感,检出限为0.04 ng·mL(-1),对其他干扰物质具有良好的选择性。电活性纳米探针可以大量制备,易于固定在SPCE上进行目标检测,并用磁铁快速更新。所提出的免疫传感器具有快速,简单,灵敏,稳定,磁体控制,无毒,无标记和可重现性。

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