LC-SRM-Based Targeted Quantification of Urinary Protein Biomarkers

Methods Mol Biol. 2018:1788:145-156. doi: 10.1007/7651_2017_93.

Abstract

Liquid chromatography (LC)-selected reaction monitoring (SRM) is a powerful protein quantification technique in terms of sensitivity, reproducibility, and multiplexing capability. LC-SRM can accurately measure the concentrations of surrogate proteotypic peptides for targeted proteins in complex biological samples by using their stable heavy isotope-labeled counterparts as internal standards. Herein, we describe a step-by-step protocol of the application of LC-SRM to quantify candidate protein biomarkers in human urine.

Keywords: Biomarker; LC-SRM; Skyline; Stable heavy isotope-labeled peptide; Targeted quantification; Urine.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Biomarkers / analysis
  • Biomarkers / urine
  • Chromatography, Liquid / methods*
  • Humans
  • Isotope Labeling / methods
  • Peptides / analysis
  • Peptides / urine
  • Proteins / analysis*
  • Proteinuria / urine*
  • Proteomics / methods*
  • Urinalysis / methods
  • Urine Specimen Collection / methods

Substances

  • Biomarkers
  • Peptides
  • Proteins