Extending the Depth of Human Plasma Proteome Coverage Using Simple Fractionation Techniques

• DOI: 10.1021/acs.jproteome.0c00670
• Publication Date: 2021-01-20T19:35:37Z
• Authors: Gurjeet Kaur; Anne Poljak; Syed Azmal Ali; Ling Zhong; Mark J. Raftery; Perminder Sachdev
• Keywords: Proteome; Sample Preparation; Sodium dodecyl sulfate; Sample complexity

Human plasma is one of the most widely used tissues in clinical analysis, and plasma-based biomarkers are for monitoring patient health status and/or response to medical treatment avoid unnecessary invasive biopsy. Data-driven proteomics has suffered from a lack throughput detection sensitivity, largely due complexity proteome particular enormous quantitative dynamic range, estimated be between 9 13 orders magnitude lowest highest abundance protein. A major challenge identify workflows that can achieve depth coverage while minimizing sample workup maximizing throughput. In this study, we have performed intensive depletion high-abundant proteins or enrichment low-abundant using Agilent multiple affinity removal liquid chromatography (LC) column—Human 6 (Hu6), LC 14 (Hu14), ProteoMiner followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) C18 prefractionation techniques. We compared performance each these fractionation approaches method satisfies requirements analysis samples include good combination with reasonable output. report one-dimensional (1D) gel-based allows parallel processing no loss coverage, serial chromatographic separation, significantly accelerates time, particularly important large projects. Furthermore, show variety methodologies similarly high allowing flexibility selection based on project-specific needs. These considerations effort accelerate research so as provide efficient, reliable, accurate diagnoses, population-based screening, studies, other work.