Differential ultracentrifugation enables deep plasma proteomics through enrichment of extracellular vesicles. Issue 7 (27th November 2022)

• Record Type: Journal Article
• Authors: Kverneland, Anders H.; Østergaard, Ole; Emdal, Kristina Bennet; Svane, Inge Marie; Olsen, Jesper Velgaard
• Other Names: Guo Tiannan guestEditor.; Aebersold Ruedi guestEditor.
• Is Part Of: Proteomics. Volume 23:Issue 7/8(2023)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-11-27
• Subjects: biomarkers -- cancer immune therapy -- extracellular vesicles -- plasma proteomics; Proteins -- Separation -- Periodicals; Bioinformatics -- Periodicals; Proteomics -- Periodicals; Genomes -- Periodicals; Molecular genetics -- Periodicals; 572.605
• DOI: 10.1002/pmic.202200039
• Languages: English
• ISSNs: 1615-9853

Abstract

Abstract: Human plasma is a rich source of biomedical information and biomarkers. However, the enormous dynamic range of plasma proteins limits its accessibility to mass spectrometric (MS) analysis. Here, we show that enrichment of extracellular vesicles (EVs) by ultracentrifugation increases plasma proteome depth by an order of magnitude. With this approach, more than two thousand proteins are routinely and reproducibly quantified by label‐free quantification and data independent acquisition (DIA) in single‐shot liquid chromatography tandem mass spectrometry runs of less than one hour. We present an optimized plasma proteomics workflow that enables high‐throughput with very short chromatographic gradients analyzing hundred samples per day with deep proteome coverage, especially when including a study‐specific spectral library generated by repeated injection and gas‐phase fractionation of pooled samples. Finally, we test the workflow on clinical biobank samples from malignant melanoma patients in immunotherapy to demonstrate the improved proteome coverage supporting the potential for future biomarker discovery.