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Quantitative proteome profiling of normal human circulating microparticles

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  3. 2019 European League Against Rheumatism/American College of Rheumatology classification criteria for systemic lupus erythematosus

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  • Ole Østergaard
  • Christoffer T Nielsen
  • Line V Iversen
  • Søren Jacobsen
  • Julia T Tanassi
  • Niels H H Heegaard
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Circulating microparticles (MPs) are produced as part of normal physiology. Their numbers, origin, and composition change in pathology. Despite this, the normal MP proteome has not yet been characterized with standardized high-resolution methods. We here quantitatively profile the normal MP proteome using nano-LC-MS/MS on an LTQ-Orbitrap with optimized sample collection, preparation, and analysis of 12 different normal samples. Analytical and procedural variation were estimated in triply processed samples analyzed in triplicate from two different donors. Label-free quantitation was validated by the correlation of cytoskeletal protein intensities with MP numbers obtained by flow cytometry. Finally, the validity of using pooled samples was evaluated using overlap protein identification numbers and multivariate data analysis. Using conservative parameters, 536 different unique proteins were quantitated. Of these, 334 (63%) were present in all samples and represent an MP core proteome. Technical triplicates showed
Original languageEnglish
JournalJournal of Proteome Research
Volume11
Issue number4
Pages (from-to)2154-63
Number of pages10
ISSN1535-3893
DOIs
Publication statusPublished - 2012

    Research areas

  • Blood Proteins, Cell-Derived Microparticles, Female, Humans, Linear Models, Male, Principal Component Analysis, Proteome, Proteomics, Reproducibility of Results

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