Comparison of highly sensitive, multiplex immunoassay platforms for streamlined clinical cytokine quantification

Selecting the optimal platforms to quantitate cytokines is challenging due to varying performance and the plethora of options available. To compare performance of three highly sensitive, multiplex assays on three different platforms – MSD S-plex, Olink Target 48, and Quanterix SP-X – to MSD V-plex which is widely used for quantitative cytokine assay. Serum and stimulated plasma samples were analyzed across each platform. The proportion of quantifiable samples was compared for each analyte and correlation analyses were performed to relate the data. For MSD S-plex, parallelism and antibody pair knockdown experiments gauged specificity of the kit. MSD S-plex was the most sensitive multiplex platform followed by Olink Target 48, Quanterix SP-X, and MSD V-plex. Concentrations across platforms differed greatly for some cytokines, but all platforms showed strong correlation. Results for MSD S-plex were confirmed by parallelism and knockdown. MSD S-plex should be a priority platform for ultra-sensitive assay. Olink Target 48 offers an enticing combination of sensitivity and multiplex capability that warrants consideration when many cytokines require quantitation. MSD V-plex, MSD S-plex and Olink quantitative assays offer high utility across drug development programs, but fit-for-purpose performance should be assessed on a per-analyte basis.由于检测性能存在差异且可选平台繁多，筛选用于细胞因子定量的最优平台颇具挑战。本研究旨在对比三种搭载于不同平台的高灵敏度多重检测技术——MSD S-plex、Olink Target 48与Quanterix SP-X——与目前广泛应用于细胞因子定量检测的MSD V-plex的性能表现。研究对血清样本及经刺激处理的血浆样本在各平台上开展了检测分析。针对每种分析物，对比了可定量样本的占比，并通过相关性分析关联各平台的检测数据。针对MSD S-plex，研究通过平行性实验与抗体对敲降实验评估了该试剂盒的特异性。检测灵敏度排序为MSD S-plex > Olink Target 48 > Quanterix SP-X > MSD V-plex。部分细胞因子在不同平台间的检测浓度差异显著，但所有平台的检测结果均呈现较强相关性。MSD S-plex的检测结果通过平行性实验与敲降实验得到了验证。MSD S-plex可作为超灵敏检测的首选平台。Olink Target 48兼具优异灵敏度与多重检测能力，在需同时定量多种细胞因子的场景中值得优先考虑。MSD V-plex、MSD S-plex及Olink定量检测技术在药物开发流程中均具备较高应用价值，但需针对每种分析物评估其适配性表现。