Institute of Macromolecular Compounds, Russian Academy of Sciences, Bolshoy pr. 31, 199004 St. Petersburg, Russia.
Institute of Experimental Medicine, Academician Pavlov's str. 12, 197376 St. Petersburg, Russia.
Sensors (Basel). 2020 May 10;20(9):2719. doi: 10.3390/s20092719.
The early diagnostics of hepatitis C virus (HCV) infections is currently one of the most highly demanded medical tasks. This study is devoted to the development of biochips (microarrays) that can be applied for the detection of HCV. The analytical platforms of suggested devices were based on macroporous poly(glycidyl methacrylate--di(ethylene glycol) dimethacrylate) monolithic material. The biochips were obtained by the covalent immobilization of specific probes spotted onto the surface of macroporous monolithic platforms. Using the developed biochips, different variants of bioassay were investigated. This study was carried out using hepatitis C virus-mimetic particles (VMPs) representing polymer nanoparticles with a size close to HCV and bearing surface virus antigen (E2 protein). At the first step, the main parameters of bioassay were optimized. Additionally, the dissociation constants were calculated for the pairs "ligand-receptor" and "antigen-antibody" formed at the surface of biochips. As a result of this study, the analysis of VMPs in model buffer solution and human blood plasma was carried out in a format of direct and "sandwich" approaches. It was found that bioassay efficacy appeared to be similar for both the model medium and real biological fluid. Finally, limit of detection (LOD), limit of quantification (LOQ), spot-to-spot and biochip-to-biochip reproducibility for the developed systems were evaluated.
丙型肝炎病毒 (HCV) 感染的早期诊断是目前医学上需求最高的任务之一。本研究致力于开发可用于检测 HCV 的生物芯片（微阵列）。所提出设备的分析平台基于大孔聚（甲基丙烯酸缩水甘油酯-二（乙二醇）二甲基丙烯酸酯）整体材料。生物芯片通过将特异性探针共价固定在大孔整体平台表面上获得。使用开发的生物芯片，研究了不同变体的生物测定法。本研究使用丙型肝炎病毒模拟粒子 (VMP) 进行，VMP 代表尺寸接近 HCV 的聚合物纳米粒子，并带有表面病毒抗原（E2 蛋白）。在第一步中，优化了生物测定的主要参数。此外，还计算了在生物芯片表面形成的“配体-受体”和“抗原-抗体”对的解离常数。作为这项研究的结果，在直接和“夹心”方法的格式中对模型缓冲溶液和人血浆中的 VMP 进行了分析。结果表明，对于模型介质和真实生物流体，生物测定的功效似乎相似。最后，评估了所开发系统的检测限 (LOD)、定量限 (LOQ)、斑点间和生物芯片间重现性。
