Khristunova Ekaterina, Barek Jiri, Kratochvil Bohumil, Korotkova Elena, Dorozhko Elena, Vyskocil Vlastimil
National Research Tomsk Polytechnic University, Lenin Avenue 30, 634050 Tomsk, Russia; Charles University, Faculty of Science, Department of Analytical Chemistry, UNESCO Laboratory of Environmental Electrochemistry, Albertov 6, 12843 Prague 2, Czech Republic; Department of Solid State Chemistry, University of Chemistry and Technology, Prague, Technicka 5, 16628 Prague 6, Czech Republic.
National Research Tomsk Polytechnic University, Lenin Avenue 30, 634050 Tomsk, Russia; Charles University, Faculty of Science, Department of Analytical Chemistry, UNESCO Laboratory of Environmental Electrochemistry, Albertov 6, 12843 Prague 2, Czech Republic.
Bioelectrochemistry. 2020 Oct;135:107576. doi: 10.1016/j.bioelechem.2020.107576. Epub 2020 May 23.
This work reports for the first time a significantly improved and simplified electrochemical immunoassay to detect antibodies to tick-borne encephalitis virus (TBEV) using a 96-well microtiter plate as a platform for immobilization and silver nanoparticles (AgNPs) as electrochemical labels. The electrochemical assay is performed by detecting the elemental silver oxidation signal where the electroactive signalling silver species are released from the bioconjugates (Ab@AgNP, Ab@AgNP, and ProteinA@AgNP). For this purpose, AgNPs were synthesized and further tagged with biomolecules (antibodies to TBEV, cleaved antibodies to TBEV, and protein A). Signal is read by linear sweep anodic stripping voltammetry (LSASV) of silver ions (through the electrochemical stripping of accumulated elemental silver) on a graphite electrode (GE). Ab@AgNP was chosen as the best option for the new electrochemical immunoassay. The results of electrochemical measurements demonstrated that voltammetric signal increased with the increasing concentration of target antibodies to TBEV within the range from 100 to 1600 IU mL, with a detection limit of 90 IU mL. To verify the practical application of the novel electrochemical immunosensor, the quantity of immunoglobulins against TBEV in human serum was checked. The results may contribute to the development of alternative methods for monitoring TBEV in biological fluids.
这项工作首次报道了一种显著改进和简化的电化学免疫分析方法，该方法以96孔微量滴定板作为固定平台，以银纳米颗粒（AgNPs）作为电化学标记物，用于检测蜱传脑炎病毒（TBEV）抗体。电化学分析通过检测元素银的氧化信号来进行，其中电活性信号银物种从生物共轭物（Ab@AgNP、Ab@AgNP和ProteinA@AgNP）中释放出来。为此，合成了AgNPs，并进一步用生物分子（抗TBEV抗体、裂解的抗TBEV抗体和蛋白A）进行标记。通过在石墨电极（GE）上对银离子进行线性扫描阳极溶出伏安法（LSASV）（通过对积累的元素银进行电化学溶出）来读取信号。Ab@AgNP被选为新的电化学免疫分析的最佳选择。电化学测量结果表明，在100至1600 IU/mL范围内，伏安信号随目标抗TBEV抗体浓度的增加而增加，检测限为90 IU/mL。为了验证新型电化学免疫传感器的实际应用，检测了人血清中抗TBEV免疫球蛋白的含量。这些结果可能有助于开发监测生物体液中TBEV的替代方法。
