Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Electroanalytical Chemistry Laboratory, Department of Chemistry, Faculty of Sciences, Azarbaijan Shahid Madani University, Tabriz 53714-161, Iran.
Int J Biol Macromol. 2018 Dec;120(Pt B):2048-2064. doi: 10.1016/j.ijbiomac.2018.09.208. Epub 2018 Oct 1.
CA 125 is a Mucin glycoprotein and its concentration in human serum correlates with a woman's risk of developing ovarian cancer and also indicates response to therapy in diagnosed patients. Accurate detection of this large, complex protein in patient biofluids is of great clinical relevance. In this work, an innovative immunoassay for quantitation of CA 125 based on signal amplification strategy was proposed. In this work, Horseradish peroxidase (HRP) labeled CA 125-antibody (anti-CA 125) was immobilized onto a green and biocompatible nanocomposite containing poly cetyl trimethyl ammonium bromide (P (CTAB) as conductive matrix, chitosan (CS) as biocompatible agent and sliver nanoparticles (Ag NPs) as signal amplification element. Therefore, a novel nanocomposite film based P (CTAB-CS) and Ag NPs was exploited to develop a highly sensitive immunosensor for detection of CA 125 protein. Importantly, Ag NPs prepared by electrodeposition method which lead to compact morphology. Fully electrochemical methodology was used to prepare a new transducer on a glassy carbon surface which provided a high surface area to immobilize a high amount of the anti-CA 125. The surface morphology of electrode was characterized by high-resolution field emission scanning electron microscope (FE-SEM) and energy dispersive spectroscopy (EDS). The immunosensor was employed for the detection of CA 125 using cyclic voltammetry (CV) and differential pulse voltammetry (DPVs) techniques. Under optimized condition the calibration curve for CA 125 concentration by SWV and DPV was linear in 0.01-400 U/mL with lower limit of quantification of 0.001 U/mL. The method was successfully applied assay of the CA 125 in unprocessed human plasma samples.
CA125 是一种黏蛋白糖蛋白,其在人血清中的浓度与女性患卵巢癌的风险相关,也可指示已确诊患者的治疗反应。准确检测患者生物体液中的这种大型复杂蛋白质具有重要的临床意义。在本工作中,提出了一种基于信号放大策略的 CA125 定量免疫分析新方法。在本工作中,辣根过氧化物酶(HRP)标记的 CA125 抗体(抗 CA125)固定在含有十六烷基三甲基溴化铵(P(CTAB)的绿色生物相容纳米复合材料上,壳聚糖(CS)作为生物相容剂和银纳米颗粒(Ag NPs)作为信号放大元件。因此,开发了一种基于 P(CTAB-CS)和 Ag NPs 的新型纳米复合膜,用于检测 CA125 蛋白的高灵敏度免疫传感器。重要的是,通过电沉积方法制备的 Ag NPs 导致形态紧凑。完全电化学方法用于在玻璃碳表面上制备新的传感器,该传感器提供了高表面积,可固定大量的抗 CA125。通过高分辨率场发射扫描电子显微镜(FE-SEM)和能谱(EDS)对电极的表面形貌进行了表征。使用循环伏安法(CV)和差分脉冲伏安法(DPV)技术对传感器进行了 CA125 的检测。在优化条件下,CA125 的浓度通过 SWV 和 DPV 的校准曲线在 0.01-400 U/mL 范围内呈线性,检出限为 0.001 U/mL。该方法成功应用于未经处理的人血浆样品中 CA125 的测定。