Departamento de Biofísica e Radiobiologia, Universidade Federal de Pernambuco (UFPE), Recife, PE, Brazil.
Departamento de Química Fundamental, Universidade Federal de Pernambuco (UFPE), Recife, PE, Brazil.
Colloids Surf B Biointerfaces. 2023 Jan;221:112984. doi: 10.1016/j.colsurfb.2022.112984. Epub 2022 Oct 29.
We report the development of a new nanostructured electrochemical immunosensing platform for the detection of the Zika virus envelope protein (EP-ZIKV). For this, quantum dots (QDs) were explored in combination with screen-printed carbon electrodes (SPCEs) functionalized with a conductor polymeric film, formed from 2-(1H-pyrrol-1-yl)ethanamine (Py), and anti-EP DIII ZIKV antibodies. Carboxylated CdTe QDs were synthesized, characterized by optical and structural techniques, and covalently immobilized onto the SPCE/PPy surface. Then, anti-EP ZIKV antibodies were also covalently conjugated to QDs. All stages of platform assembly were evaluated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The detection of EP-ZIKV was performed by differential pulse voltammetry (DPV). Results indicated that QDs were efficiently immobilized, and did not show oxidation, under the conditions evaluated, for at least 7 months. Anti-EP ZIKV antibodies were effectively immobilized on the PPy/QDs surface, even after 2 months of electrode storage. The platform enabled the detection of EP-ZIKV with high sensitivity using minimal sample volumes (LOD = 0.1 ng mL and LOQ = 0.4 ng mL). The platform was also able to detect EP-ZIKV in spiked serum samples. Moreover, the platform showed specificity, not detecting the EP-DENV 3 nor a mixture of four DENV serotypes antigens. Thus, the proposed combination favored the development of a sensitive immunosensing platform, promising for the detection of Zika in the viremic phase, which also holds potential for transposition to other arboviruses.
我们报告了一种新的纳米结构电化学免疫传感平台的开发,用于检测寨卡病毒包膜蛋白(EP-ZIKV)。为此,我们探索了量子点(QDs)与功能化的丝网印刷碳电极(SPCE)结合使用的可能性,该功能化电极的导体聚合物膜由 2-(1H-吡咯-1-基)乙胺(Py)和抗 EP DIII ZIKV 抗体组成。合成了羧基化 CdTe QDs,通过光学和结构技术进行了表征,并将其共价固定在 SPCE/PPy 表面上。然后,还将抗 EP ZIKV 抗体共价偶联到 QDs 上。通过循环伏安法(CV)和电化学阻抗谱(EIS)评估了平台组装的所有阶段。通过差分脉冲伏安法(DPV)进行 EP-ZIKV 的检测。结果表明,在评估的条件下,QDs 可以有效固定,并且至少在 7 个月内不会发生氧化。抗 EP ZIKV 抗体即使在电极储存 2 个月后也可以有效固定在 PPy/QDs 表面上。该平台使用最小的样品量(LOD = 0.1ng/mL 和 LOQ = 0.4ng/mL)实现了对 EP-ZIKV 的高灵敏度检测。该平台还能够检测到加标血清样本中的 EP-ZIKV。此外,该平台表现出特异性,不会检测到 EP-DENV 3 或四种 DENV 血清型抗原的混合物。因此,所提出的组合有利于开发敏感的免疫传感平台,有望用于检测病毒血症期的寨卡病毒,也有可能转移到其他虫媒病毒。