Multiplexed panel of precisely quantified salivary proteins for biomarker assessment

Andrew J. Percy, Darryl B. Hardie, Armando Jardim, Juncong Yang, Monica H. Elliott, Suping Zhang, Yassene Mohammed, Christoph H. Borchers

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

An increasingly popular “absolute” quantitative technique involves the SRM or MRM approach with stable isotope-labeled standards (SIS). Using this approach, many proteins in human plasma/serum have been quantified for biomarker assessment and disease stratification. Due to the complexity of plasma and the invasive nature of its collection, alternative biosamples are currently being explored. Here, we present the broadest panel of multiplexed MRM assays with SIS peptides for saliva proteins developed to date. The validated panel consists of 158 candidate human saliva protein biomarkers, inferred from 244 interference-free peptides. The resulting concentrations were reproducibly quantified over a 6 order-of-magnitude concentration range (from 218 μg/mL to 88 pg/mL; average CVs of 12% over analytical triplicates). All concentrations were determined from reverse standard curves, which were generated using a constant concentration of endogenous material with varying concentrations of spiked-in SIS peptides. The large-scale screening of the soluble and membrane-associated proteins contained within the 158-plex assay could present new opportunities for biomarker assessment and clinical diagnostics.

Original languageEnglish
Article number1600230
JournalProteomics
Volume17
Issue number6
DOIs
Publication statusPublished - 1 Mar 2017
Externally publishedYes

Keywords

  • Biomarker
  • Biomedicine
  • MRM
  • Quantitation
  • Saliva
  • Targeted

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