Ben Neely (@benneely.com)

This banger from Matt @mattwfoster.bsky.social et al. just dropped. Love some whole blood proteomics fun, and really so much of what Matt has been doing with blood. Excellent work! pubs.acs.org/doi/10.1021/...

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Development and Validation of a Streamlined Workflow for Proteomic Analysis of Proteins and Post-translational Modifications from Dried Blood It is increasingly recognized that the ‘omic analysis of whole blood has applications for precision medicine and disease phenotyping. Despite this realization, whole blood is generally viewed as a challenging analytical matrix in comparison to plasma or serum. Moreover, proteomic analyses of whole blood have almost exclusively focused on (non)targeted analyses of protein abundances and much less on post-translational modifications (PTMs). Here, we developed a streamlined workflow for processing 20 microliters of venous blood collected by volumetric absorptive microsampling that incorporates serial trypsinization and N-glycopeptide and phosphopeptide enrichment and avoids laborious sample dry-down or cleanup steps. As many as 10,000 analytes (reported as protein groups, glycopeptidoforms, and phosphosites) can be quantified by liquid chromatography-tandem mass spectrometry in under 2 h of MS acquisition time. Using these methods, we explored the stability of “dried” and “wet” blood proteomes, as well as the effects of ex vivo inflammatory stimulus or phosphatase inhibition. Multiomics factor analysis enabled facile identification of analytes that contributed to interindividual variability of the blood proteomes, including N-glycopeptides that distinguish immunoglobulin heavy constant alpha 2 allotypes. Collectively, our results help to establish feasibility and best practices for the integrated MS-based quantification of proteins and PTMs from dried blood. https://pubs.acs.org/doi/10.1021/acs.jproteome.6c00032