title: SRM/MRM targeted proteomics as a tool for biomarker validation and absolute quantification in human urine.
abstract: Urine has always been one of the most suitable body fluids for clinical applications. Absolute quantification of disease protein biomarkers in body fluids such as urine is a key step in the biomarker development pipeline. Nevertheless, identification of groups of proteins in complex biological samples is challenging. Traditional affinity-based methodologies such as ELISA are used to verify the presence of biomarkers in clinical samples. More recently, targeted mass spectrometry-based strategies have been developed for biomarker validation, offering an alternative. The great advantage of targeted mass spectrometry-based methodologies is that they allow accurate and specific simultaneous quantification of several biomarkers (multiplexing). Peptides are used as protein surrogates, measured using triple quadrupole instruments in selected reaction monitoring/multiple reaction monitoring mode. In this review, the workflow of selected reaction monitoring/multiple reaction monitoring for disease biomarker validation in urine is presented and assay performance in the latest studies is described.
authors.forename: George
authors.initials: G
authors.lastname: Mermelekas
authors.affiliation: a Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Greece Soranou Efessiou 4, 11527, Athens, Greece.
authors.forename: Antonia
authors.initials: A
authors.lastname: Vlahou
authors.affiliation: a Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Greece Soranou Efessiou 4, 11527, Athens, Greece.
authors.forename: Jerome
authors.initials: J
authors.lastname: Zoidakis
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000652:urine*; D003627:Data Interpretation, Statistical; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D057188:Workflow
chemical_list: D015415:Biomarkers; D020543:Proteome
keywords: absolute quantification; biomarker validation; multiple reaction monitoring; selected reaction monitoring; targeted proteomics; urine