Chair of Analytical Chemistry and Water Chemistry, TUM School of Natural Sciences, Technical University of Munich, Lichtenbergstr. 4, 85748 Garching, Germany.
Center of Allergy and Environment (ZAUM), Technical University of Munich and Helmholtz Center Munich, Member of the German Center of Lung Research (DZL), 80802 Munich, Germany.
Biosensors (Basel). 2023 Jun 22;13(7):670. doi: 10.3390/bios13070670.
Protein biomarkers in nasal secretions can be used as a measure to differentiate between allergies, airway diseases and infections for non-invasive diagnostics. The point-of-care quantification of biomarker levels using flow-based microarray facilitates precise and rapid diagnosis and displays the potential for targeted and effective treatment. For the first time, we developed a flow-based chemiluminescence sandwich microarray immunoassay (CL-SMIA) for the quantification of nasal interferon-beta (IFN-β) on the Microarray Chip Reader-Research (MCR-R). Polycarbonate foils are used as a cost-effective surface for immobilizing capture antibodies. By using a commercially available set of anti-human IFN-β antibodies, the CL-SMIA can be compared directly to an enzyme-linked immunosorbent assay (ELISA) performed in microtiter plates concerning the bioanalytical performance and economic issues. Pre-incubation of the sample with detection antibodies facilitates the lower consumption of detection antibodies, as this allows for a longer interaction time between the antibody and the biomarker. The direct injection of pre-incubated samples into the microarray chips eliminates the adsorption of proteins in the tubing as well as the contamination of the tubing and valves of the MCR-R with clinical samples. The small flow cell allows for a low sample volume of 50 μL. The limit of detection of 4.53 pg mL was slightly increased compared to a sandwich ELISA performed on microtiter plates which were 1.60 pg mL. The possibility to perform the CL-SMIA in a multiplexed mode makes it a promising assay for the rapid and cost-effective non-invasive detection of biomarkers in nasal secretions.
鼻腔分泌物中的蛋白质生物标志物可用作区分过敏、气道疾病和感染的指标,以实现非侵入性诊断。使用基于流动的微阵列进行即时护理点定量生物标志物水平有助于进行精确和快速诊断,并显示出针对目标和有效治疗的潜力。我们首次在 Microarray Chip Reader-Research (MCR-R) 上开发了基于流动的化学发光夹心微阵列免疫分析(CL-SMIA),用于定量鼻腔干扰素-β(IFN-β)。聚碳酸酯箔可用作固定捕获抗体的经济有效的表面。通过使用市售的一组抗人 IFN-β 抗体,CL-SMIA 可以直接与微滴定板上进行的酶联免疫吸附测定(ELISA)进行比较,涉及生物分析性能和经济问题。在检测抗体中预孵育样品可以减少检测抗体的消耗,因为这允许抗体和生物标志物之间有更长的相互作用时间。将预孵育的样品直接注入微阵列芯片可以消除蛋白质在管中的吸附,以及临床样品对 MCR-R 的管和阀门的污染。小的流通池允许样品体积低至 50 μL。与在微滴定板上进行的夹心 ELISA 相比,检测限略微增加到 4.53 pg mL,为 1.60 pg mL。以多重模式进行 CL-SMIA 的可能性使其成为一种有前途的分析方法,可用于快速、经济高效地非侵入性检测鼻腔分泌物中的生物标志物。