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一种用于人血清中神经元特异性烯醇化酶检测的高选择性聚(噻吩)接枝聚(甲基丙烯酰胺)聚合物修饰 ITO 电极。

A Highly Selective Poly(thiophene)-graft-Poly(methacrylamide) Polymer Modified ITO Electrode for Neuron Specific Enolase Detection in Human Serum.

机构信息

Tekirdagˇ Namık Kemal University, Scientific and Technological Research Center, Tekirdagˇ, 59000, Turkey.

Çanakkale Onsekiz Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, 17000, Turkey.

出版信息

Macromol Biosci. 2019 Aug;19(8):e1900109. doi: 10.1002/mabi.201900109. Epub 2019 Jun 21.

Abstract

In this study, an impedimetric immunosensor based on polymer poly(thiophene)-graft-poly(methacrylamide) polymer (P(Thi-g-MAm)) modified indium tin oxide (ITO) electrode is developed for the detection of the Neuron Specific Enolase (NSE) cancer biomarker. First, the P(Thi-g-MAm) polymer is synthesized and coated on the ITO electrode by using a spin-coating technique. P(Thi-g-MAm) polymer acts as an immobilization platform for immobilization of NSE-specific monoclonal antibodies. Anti-NSE antibodies are utilized as biosensing molecules and they bind to the amino groups of P(Thi-g-Mam) polymer via glutaraldehyde cross-linking. Spin-coating technique is employed for bioelectrode fabrication and this technique provides a thin and uniform film on the ITO electrode surface. This bioelectrode fabrication technique is simple and it generates a suitable platform for large-scale loadings of anti-NSE antibodies. This immunosensor exhibits a wide linear detection range from 0.02 to 4 pg mL and with an ultralow detection limit of 6.1 fg mL . It reveals a good long-term stability (after 8 weeks, 78% of its initial activity), an excellent reproducibility (1.29% of relative standard deviation (RSD)), a good repeatability (5.55% of RSD), and a high selectivity. In addition, the developed immunosensor is proposed as a robust diagnostic tool for the clinical detection of NSE and other cancer biomarkers.

摘要

在这项研究中,开发了一种基于聚合物聚(噻吩)-接枝-聚(甲基丙烯酰胺)(P(Thi-g-MAm))修饰的氧化铟锡(ITO)电极的阻抗免疫传感器,用于检测神经元特异性烯醇化酶(NSE)癌症生物标志物。首先,通过旋涂技术合成 P(Thi-g-MAm)聚合物并将其涂覆在 ITO 电极上。P(Thi-g-MAm)聚合物作为固定化平台,用于固定 NSE 特异性单克隆抗体。抗 NSE 抗体用作生物传感分子,它们通过戊二醛交联与 P(Thi-g-Mam)聚合物的氨基结合。旋涂技术用于生物电极的制造,这种技术在 ITO 电极表面提供了一层薄而均匀的薄膜。这种生物电极制造技术简单,为大量负载抗 NSE 抗体提供了合适的平台。该免疫传感器的线性检测范围从 0.02 到 4 pg mL,检测限低至 6.1 fg mL。它具有良好的长期稳定性(8 周后,其初始活性的 78%)、优异的重现性(相对标准偏差为 1.29%)、良好的重复性(相对标准偏差为 5.55%)和高选择性。此外,所开发的免疫传感器被提议作为一种用于临床检测 NSE 和其他癌症生物标志物的强大诊断工具。

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