Departments of Biological Sciences, University of Alberta, Edmonton, AB, T6G 2E8, Canada.
Department of Electrical and Computer Engineering, Donadeo Innovation Centre for Engineering, University of Alberta, 11-358, 9211-116 St, Edmonton, AB, T6G 2H5, Canada.
Sci Rep. 2022 Jul 27;12(1):12850. doi: 10.1038/s41598-022-17219-7.
As COVID-19 waves continue to spread worldwide, demand for a portable, inexpensive and convenient biosensor to determine community immune/infection status is increasing. Here we describe an impedance-based affinity biosensor using Interdigitated Electrode (IDE) arrays to detect antibodies to SARS-CoV-2 in serum. We created the biosensor by functionalizing the IDEs' surface with abaculaovirus-expressed and purified Spike (S) protein to bind anti-SARS CoV-2antibodies. Gold nanoparticles (GNP) fused to protein G were used to probe for bound antibodies. An ELISA assay using horseradish peroxidase-protein G to probe for bound IgG confirmed that the purified S protein bound a commercial source of anti-SARS-CoV-2 antibodies specifically and bound anti-SARS-CoV-2 antibodies in COVID-19 positive serum. Then we demonstrated that our biosensor could detect anti-SARS-CoV-2 antibodies with 72% sensitivity in 2 h. Using GNP-protein G, the affinity biosensor had increased impedance changes with COVID-19positive serum and minimal or decreased impedance changes with negative serum. This demonstrated that our biosensor could discriminate between COVID-19 positive and negative sera, which were further improved using poly(vinyl alcohol)as a blocking agent.
随着 COVID-19 浪潮在全球范围内持续蔓延,人们对便携式、廉价且方便的生物传感器的需求越来越大,以便确定社区的免疫/感染状态。在这里,我们描述了一种基于阻抗的亲和生物传感器,该传感器使用叉指电极 (IDE) 阵列来检测血清中的 SARS-CoV-2 抗体。我们通过用杆状病毒表达和纯化的 Spike (S) 蛋白功能化 IDE 的表面来创建生物传感器,以结合抗 SARS-CoV-2 抗体。与蛋白 G 融合的金纳米颗粒 (GNP) 用于探测结合的抗体。使用辣根过氧化物酶-蛋白 G 进行 ELISA 检测以探测结合的 IgG 证实,纯化的 S 蛋白特异性结合商业来源的抗 SARS-CoV-2 抗体,并结合 COVID-19 阳性血清中的抗 SARS-CoV-2 抗体。然后,我们证明我们的生物传感器可以在 2 小时内以 72%的灵敏度检测抗 SARS-CoV-2 抗体。使用 GNP-蛋白 G,亲和生物传感器对 COVID-19 阳性血清的阻抗变化增加,而对阴性血清的阻抗变化最小或减少。这表明我们的生物传感器可以区分 COVID-19 阳性和阴性血清,进一步使用聚乙烯醇作为阻断剂得到了改善。