Laboratório de Sensores Eletroquímicos (LabSensE), Departamento de Química, Universidade Federal Do Paraná (UFPR), CP 19032, CEP, 81531-990, Curitiba, PR, Brazil.
Departamento de Ciências, Matemática e Educação. Universidade Federal de São Carlos (UFSCar), 13600-970, Araras, SP, Brazil.
Anal Chim Acta. 2021 Feb 22;1147:30-37. doi: 10.1016/j.aca.2020.12.014. Epub 2020 Dec 11.
Simple, low-cost, and sensitive new platforms for electrochemical immunosensors for virus detection have been attracted attention due to the recent pandemic caused by a new type of coronavirus (SARS-CoV-2). In the present work, we report for the first time the construction of an immunosensor using a commercial 3D conductive filament of carbon black and polylactic acid (PLA) to detect Hantavirus Araucaria nucleoprotein (Np) as a proof-of-concept. The recognition biomolecule was anchored directly at the filament surface by using N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-Hydroxysuccinimide (EDC/NHS). Conductive and non-conductive composites of PLA were characterized using thermal gravimetric analysis (TGA), revealing around 30% w/w of carbon in the filament. Morphological features of composites were obtained from SEM and TEM measurements. FTIR measurement revealed that crosslinking agents were covalently bonded at the filament surface. Electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for the evaluation of each step involved in the construction of the proposed immunosensor. The results showed the potentiality of the device for the quantitative detection of Hantavirus Araucaria nucleoprotein (Np) from 30 μg mL to 240 μg mL with a limit of detection of 22 μg mL. Also, the proposed immunosensor was applied with success for virus detection in 100x diluted human serum samples. Therefore, the PLA conductive filament with carbon black is a simple and excellent platform for immunosensing, which offers naturally carboxylic groups able to anchor covalently biomolecules.
由于新型冠状病毒（SARS-CoV-2）引起的最近一次大流行，用于病毒检测的简单、低成本和灵敏的新型电化学免疫传感器平台引起了人们的关注。在本工作中，我们首次报道了使用商业 3D 碳黑和聚乳酸（PLA）导电长丝构建免疫传感器来检测汉坦病毒 Araucaria 核蛋白（Np）的情况，作为概念验证。通过使用 N-（3-二甲基氨基丙基）-N'-乙基碳二亚胺盐酸盐和 N-羟基琥珀酰亚胺（EDC/NHS），将识别生物分子直接固定在长丝表面。使用热重分析（TGA）对 PLA 的导电和非导电复合材料进行了表征，发现长丝中含有约 30wt%的碳。通过 SEM 和 TEM 测量获得了复合材料的形貌特征。FTIR 测量表明交联剂被共价键合在长丝表面。使用循环伏安法（CV）和电化学阻抗谱（EIS）等电化学技术对构建所提出的免疫传感器涉及的每一步进行了评估。结果表明，该器件具有从 30μg mL 到 240μg mL 定量检测汉坦病毒 Araucaria 核蛋白（Np）的潜力，检测限为 22μg mL。此外，该免疫传感器还成功地应用于 100 倍稀释的人血清样品中的病毒检测。因此，碳黑填充 PLA 导电长丝是一种简单而出色的免疫传感平台，它提供了能够共价固定生物分子的天然羧酸基团。
