Abstract
Proteins in the circulatory system mirror an individual's physiology. In daily clinical practice, protein levels are generally determined using single-protein immunoassays. High-throughput, quantitative analysis using mass-spectrometry-based proteomics of blood, plasma, and serum would be advantageous but is challenging because of the high dynamic range of protein abundances. Here, we introduce a rapid and robust "plasma proteome profiling" pipeline. This single-run shotgun proteomic workflow does not require protein depletion and enables quantitative analysis of hundreds of plasma proteomes from 1μl single finger pricks with 20min gradients. The apolipoprotein family, inflammatory markers such as C-reactive protein, gender-related proteins, and >40 FDA-approved biomarkers are reproduciblyquantified (CV <20% with label-free quantification). Furthermore, we functionally interpret a 1,000-protein,quantitative plasma proteome obtained by simple peptide pre-fractionation. Plasma proteome profiling delivers an informative portrait of a person's health state, and we envision its large-scale use in biomedicine.
Highlights
Blood, plasma, and serum are the predominant samples used for diagnostic analyses in clinical practice and are available in biobanks from thousands of clinical studies (Vegvari et al, 2011)
Mass spectrometry (MS)based proteomics is a technology that could address all of these limitations and that should be capable of discovering biomarkers in this accessible body fluid (Anderson, 2014)
MS-based plasma proteomics is extremely challenging for a number of reasons, most prominently the extremely large dynamic range of protein abundances (Anderson and Anderson, 2002; Omenn, 2005)
Summary
Plasma, and serum are the predominant samples used for diagnostic analyses in clinical practice and are available in biobanks from thousands of clinical studies (Vegvari et al, 2011). Immunoassays have inherent limitations with regard to multiplexing, their specificity for protein isoforms, and their incompatibility with hypothesis-free investigations. Mass spectrometry (MS)based proteomics is a technology that could address all of these limitations and that should be capable of discovering biomarkers in this accessible body fluid (Anderson, 2014). MS-based plasma proteomics is extremely challenging for a number of reasons, most prominently the extremely large dynamic range of protein abundances (Anderson and Anderson, 2002; Omenn, 2005). Only few novel biomarkers have been established—fewer than 1.5 per year in the 15 years before 2010 (Anderson, 2010)— and this has generally been done by immunoassay-based technologies, such as prostate-specific antigen, one of the best known biomarkers in medicine (Vihko et al, 1978)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
