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Proteomics. 2017 Mar;17(6). doi: 10.1002/pmic.201600230. Epub 2016 Oct 19.

Multiplexed panel of precisely quantified salivary proteins for biomarker assessment.

Author information

1
University of Victoria-Genome British Columbia Proteomics Centre, Vancouver Island Technology Park, Victoria, BC, Canada.
2
Institute of Parasitology, McGill University, Montreal, QC, Canada.
3
MRM Proteomics, Vancouver Island Technology Park, Victoria, BC, Canada.
4
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands.
5
Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.

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.

KEYWORDS:

Biomarker; Biomedicine; MRM; Quantitation; Saliva; Targeted

PMID:
27538354
DOI:
10.1002/pmic.201600230
[Indexed for MEDLINE]

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