Towards the Development of a 3-D Biochip for the Detection of Hepatitis C Virus

Abstract

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-co-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.

Molecularly imprinted macroporous polymer monolithic layers for L‐phenylalanine recognition in complex biological fluids

“…Silanization of supporting glass slides A well (25 × 18 × 0.2 mm) on a glass slide, preliminary fabricated as described elsewhere, 45 was filled with 200 μl of 15% solution of 3-(trimethoxysilyl)propyl methacrylate in toluene (v/v) and left for 12 h in a dark at 22 C. After that, the glass slide was washed 3 times with acetone and ethanol, respectively, and dried in a dark at 40 C.…”

Nanomaterials for virus sensing and tracking

“…446 Polylactic acid-polyethylene glycol copolymer nanoparticles (73 AE 11 nm) with E2 surface functionalization possess virus mimetic properties and are essential for the development of HCV-sensitive 3D biochips. 423 Polyethyleneimine functionalized with polystyrene nanospheres (120 nm) via coordination polymerization of ferrocene derivatives exhibit a singular morphology as shown in Fig. 20A with a tunable, biocompatible structure.…”

Macroporous Polymer Monoliths in Thin Layer Format

“…Another practical application of monolithic-based microarray was their application for the detection of the E2 protein and hepatitis C virus-mimicking nanoparticles. The applicability of poly(GMA-co-DEGDMA) layers with different pore size as microarray platforms for direct and "sandwich" protocols based on binding of VMPs to immobilized anti-E2 antibodies or recombinant CD81-LEL was testified [114]. The identical detection efficiency of VMPs in a model buffer medium and spiked human blood plasma has proven the suitability of discussed materials for analysis in real biological fluids.…”