Science for Life Laboratory, Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, Sweden.
Department of Clinical Medicine, Faculty of Health Science, The Arctic University of Tromsö, Tromsö, Norway.
Methods Mol Biol. 2021;2344:65-78. doi: 10.1007/978-1-0716-1562-1_5.
Protein biomarkers in biological fluids represent an important resource for improving the clinical management of diseases. Current proteomics technologies are capable of performing high-throughput and multiplex profiling in different types of fluids, often leading to the shortlisting of tens of candidate biomarkers per study. However, before reaching any clinical setting, these discoveries require thorough validation and an assay that would be suitable for routine analyses. In the path from biomarker discovery to validation, the performance of the assay implemented for the intended protein quantification is extremely critical toward achieving reliable and reproducible results. Development of robust sandwich immunoassays for individual candidates is challenging and labor and resource intensive, and multiplies when evaluating a panel of interesting candidates at the same time. Here we describe a versatile pipeline that facilitates the systematic and parallel development of multiple sandwich immunoassays using a bead-based technology.
生物体液中的蛋白质生物标志物是改善疾病临床管理的重要资源。当前的蛋白质组学技术能够在不同类型的体液中进行高通量和多重分析,通常会在每项研究中筛选出数十种候选生物标志物。然而,在进入任何临床环境之前,这些发现需要进行彻底的验证,并开发出适合常规分析的检测方法。在从生物标志物发现到验证的过程中,用于定量目标蛋白质的检测方法的性能对于获得可靠和可重复的结果至关重要。针对单个候选者开发稳健的夹心免疫测定法具有挑战性,需要大量的人力和物力,并且在同时评估一组有趣的候选者时会成倍增加。在这里,我们描述了一个通用的管道,该管道使用基于珠的技术促进了多个夹心免疫测定法的系统和并行开发。