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基于肽纳米管-金纳米粒子-聚苯胺固定化的铅笔芯石墨电极电化学免疫测定法检测前列腺特异性抗原。

Electrochemical immunoassay for detection of prostate specific antigen based on peptide nanotube-gold nanoparticle-polyaniline immobilized pencil graphite electrode.

机构信息

Biochemistry Division, Department of Chemistry, Hacettepe University, 06800 Ankara, Turkey.

Analytical Chemistry Division, Department of Chemistry, Hacettepe University, 06800 Ankara, Turkey.

出版信息

J Colloid Interface Sci. 2018 Jan 15;510:318-326. doi: 10.1016/j.jcis.2017.09.079. Epub 2017 Sep 21.

Abstract

In this work, we developed a disposable amperometric sandwich-type immunoassay to detect prostate specific antigen (PSA). A self-assembled peptide nanotube (PNT), gold nanoparticle (AuNP) and polyaniline (PANI) composite (PANI/AuNP-PNT) were used to modify a pencil graphite electrode (PGE). Anti-PSA (Ab1) was immobilized on the modified electrode (PANI/AuNP-PNT/PGE) to capture PSA. Horseradish peroxidase (HRP) labeled anti-PSA (HRP-Ab2) was used as a tracer antibody. The modified electrodes were characterized with scanning electron microscopy (SEM), thermogravimetric analysis (TGA), energy dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). PSA concentration in phosphate buffer (pH=7.4) was determined with electro-catalytic reduction of HO on the modified working electrode by using the chronoamperometric method. Limit of detection was found out to be 0.68ng/mL in a linear range of 1-100ng/mL with a high regression (R=0.990). To show the practicality of the modified biosensor in real matrixes, it was successfully applied for the detection of PSA in blood serum samples. The proposed method was also compared with enzyme-linked immunosorbent assay (ELISA) and compatible results were obtained. The developed immunoassay exhibited good reproducibility together with high stability and provides an efficient approach to detect PSA cost-effectively compared to traditional methods.

摘要

在这项工作中,我们开发了一种用于检测前列腺特异性抗原 (PSA) 的一次性安培型三明治免疫分析。自组装的肽纳米管 (PNT)、金纳米颗粒 (AuNP) 和聚苯胺 (PANI) 复合材料 (PANI/AuNP-PNT) 被用于修饰铅笔石墨电极 (PGE)。抗 PSA (Ab1) 被固定在修饰电极 (PANI/AuNP-PNT/PGE) 上以捕获 PSA。辣根过氧化物酶 (HRP) 标记的抗 PSA (HRP-Ab2) 被用作示踪抗体。修饰电极的特性包括扫描电子显微镜 (SEM)、热重分析 (TGA)、能量色散 X 射线光谱 (EDS)、透射电子显微镜 (TEM)、循环伏安法 (CV) 和电化学阻抗谱 (EIS)。通过使用计时安培法在修饰工作电极上电催化还原 HO 来测定磷酸盐缓冲液 (pH=7.4) 中的 PSA 浓度。在 1-100ng/mL 的线性范围内,检测限为 0.68ng/mL,具有较高的回归系数 (R=0.990)。为了展示修饰生物传感器在实际基质中的实用性,它成功地应用于血清样品中 PSA 的检测。该方法还与酶联免疫吸附测定 (ELISA) 进行了比较,得到了兼容的结果。与传统方法相比,所开发的免疫分析具有良好的重现性、高稳定性,并且能够经济有效地检测 PSA。

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