Tekirdagˇ Namık Kemal University, Scientific and Technological Research Center, Tekirdagˇ, 59000, Turkey.
Çanakkale Onsekiz Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, 17000, Turkey.
Macromol Biosci. 2019 Aug;19(8):e1900109. doi: 10.1002/mabi.201900109. Epub 2019 Jun 21.
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.
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。