Evaluation of Olink Reveal Proximity Extension Assay for High-Throughput Proteomics: A Case Study Using NIST SRM 1950 and Spike-In Protein Standards

Plasma proteomics has regained attention in recent years through advancements in mass spectrometry instrumentation and sample preparation, as well as new high-throughput affinity-based technologies. Here, we evaluate the analytical performance of the new Olink Reveal platform, a proximity extension assay based technology quantifying 1,034 proteins across biological pathways. Using spiked-in recombinant Interleukin-10 (IL-10) and vascular endothelial growth factor D (VEGF-D) in the NIST SRM 1950 plasma standard, we assessed the linearity, sensitivity, precision and accuracy of the Olink assay. The results demonstrated strong linear relationships (R² 0.922–0.953) for both IL-10 and VEGF-D across spiked-in concentrations, confirming the robust technical performance of Olink Reveal and underscoring its suitability for relative quantitation in large-scale studies. The resulting data contains no sensitive or personally identifiable information, and is available in public repositories, and therefore suitable for use in benchmarking and software development.

近年来，随着质谱仪器设备与样品前处理技术的进步，以及新型高通量亲和检测技术的发展，血浆蛋白质组学再度引发研究关注。本研究针对新型Olink Reveal平台——一种基于邻近延伸分析法（proximity extension assay）的检测技术——的分析性能展开评估，该平台可定量覆盖生物通路中的1034种蛋白质。本研究以NIST SRM 1950血浆标准物质为基质，向其中加入重组白细胞介素-10（Interleukin-10，IL-10）与血管内皮生长因子D（vascular endothelial growth factor D，VEGF-D），以此评估Olink检测的线性范围、灵敏度、精密度与准确度。实验结果显示，在加标浓度范围内，IL-10与VEGF-D均呈现出极佳的线性相关性（决定系数R²为0.922~0.953），证实了Olink Reveal平台稳健的技术性能，同时凸显其适用于大规模研究中的相对定量分析。本研究产生的数据不包含敏感信息或个人可识别信息，且已公开存储于公共数据库中，因此可用于基准测试与软件开发相关研究。