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Publications

Journal Article Preprint Epublish

PubMed: 40492198 DOI: 10.1101/2025.04.22.650109

Mass spectrometry-based quantification of proteins and post-translational modifications in dried blood: longitudinal sampling of patients with sepsis in Tanzania.

bioRxiv

Authors: Matthew W Foster, Timothy J McMahon, James S Ngocho, Asia H Kipengele, Marlene Violette, Youwei Chen, Deng B Madut, Robert S Plumb, A Ian Wong, Lingye Chen, Grace M Lee, Philoteus A Sakasaka, Blandina T Mmbaga, John A Crump, Micah T McClain, Christopher W Woods, Venance P Maro, Matthew P Rubach

Affiliations

  • Department of Medicine, Duke University School of Medicine, Durham NC USA.
  • Duke Proteomics and Metabolomics Core Facility, Duke University School of Medicine, Durham NC USA.
  • Medicine Service, Durham VA Healthcare System, Durham NC USA.
  • School of Medicine, Institute of Public Health, KCMC University, Moshi, Tanzania.
  • Kilimanjaro Clinical Research Institute, Kilimanjaro Christian Medical Centre, Moshi Tanzania.
  • Waters Corporation, Milford MA USA.
  • Department of Medicine, Duke University School of Medicine, Durham NC USA. Medicine Service, Durham VA Healthcare System, Durham NC USA. Department of Biostatistics and Bioinformatics, Duke University School of Medicine, Durham NC USA.
  • Duke Global Health Institute, Durham NC USA.
  • Department of Medicine, Duke University School of Medicine, Durham NC USA. Medicine Service, Durham VA Healthcare System, Durham NC USA. Duke Global Health Institute, Durham NC USA.

Abstract

The proteomic analysis of blood is routine for disease phenotyping and biomarker development. Whole blood is commonly separated into soluble and cellular fractions. However, this can introduce pre-analytical variability; and analysis of a single component (which is common) may ignore important pathophysiology. We have recently developed methods for the facile processing of dried blood for mass spectrometry-based quantification of the proteome, N-glycoproteome and phosphoproteome. Here, we applied this approach to 38 patients in Tanzania who presented to the hospital with sepsis. Blood was collected on Mitra devices at presentation and 1, 3 and 28-42 days post-enrollment. Processing of 96 total samples was performed in plate-based formats and completed within two days. Approximately 2,000 protein groups and 8,000 post-translational modifications were quantified in 3 LC-MS/MS runs at ~1.5 hours per sample. Analysis of differential abundance revealed blood proteome signatures of acute phase response and neutrophilic inflammation that partially resolved at the 28-42 day timepoint. Numerous analytes correlated with clinical laboratory values for c-reactive protein and white blood cell counts, as well as the Universal Vital Assessment illness severity score. These datasets serve as proof-of-concept for large scale MS-based (sub)phenotyping of disease using dried blood and are available via the ProteomeXchange consortium (PXD060377).

MeSH

Cells, Blood, Sepsis, Protein Processing, Post-Translational, Proteome