title: Accurate quantification of cardiovascular biomarkers in serum using Protein Standard Absolute Quantification (PSAQ™) and selected reaction monitoring.
abstract: Development of new biomarkers needs to be significantly accelerated to improve diagnostic, prognostic, and toxicity monitoring as well as therapeutic follow-up. Biomarker evaluation is the main bottleneck in this development process. Selected Reaction Monitoring (SRM) combined with stable isotope dilution has emerged as a promising option to speed this step, particularly because of its multiplexing capacities. However, analytical variabilities because of upstream sample handling or incomplete trypsin digestion still need to be resolved. In 2007, we developed the PSAQ™ method (Protein Standard Absolute Quantification), which uses full-length isotope-labeled protein standards to quantify target proteins. In the present study we used clinically validated cardiovascular biomarkers (LDH-B, CKMB, myoglobin, and troponin I) to demonstrate that the combination of PSAQ and SRM (PSAQ-SRM) allows highly accurate biomarker quantification in serum samples. A multiplex PSAQ-SRM assay was used to quantify these biomarkers in clinical samples from myocardial infarction patients. Good correlation between PSAQ-SRM and ELISA assay results was found and demonstrated the consistency between these analytical approaches. Thus, PSAQ-SRM has the capacity to improve both accuracy and reproducibility in protein analysis. This will be a major contribution to efficient biomarker development strategies.
authors.affiliation: CEA, IRTSV, Biologie à Grande Echelle, F-38054 Grenoble, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Huillet
authors.forename: Annie
authors.initials: A
authors.lastname: Adrait
authors.forename: Dorothée
authors.initials: D
authors.lastname: Lebert
authors.forename: Guillaume
authors.initials: G
authors.lastname: Picard
authors.forename: Mathieu
authors.initials: M
authors.lastname: Trauchessec
authors.forename: Mathilde
authors.initials: M
authors.lastname: Louwagie
authors.forename: Alain
authors.initials: A
authors.lastname: Dupuis
authors.forename: Luc
authors.initials: L
authors.lastname: Hittinger
authors.forename: Bijan
authors.initials: B
authors.lastname: Ghaleh
authors.forename: Philippe
authors.initials: P
authors.lastname: Le Corvoisier
authors.forename: Michel
authors.initials: M
authors.lastname: Jaquinod
authors.forename: Jérôme
authors.initials: J
authors.lastname: Garin
authors.forename: Christophe
authors.initials: C
authors.lastname: Bruley
authors.forename: Virginie
authors.initials: V
authors.lastname: Brun
mesh_terms: D015415:Biomarkers / Q000097:blood*; D001798:Blood Proteins / Q000032:analysis*; D016022:Case-Control Studies; D002853:Chromatography, Liquid; D003327:Coronary Disease / Q000097:blood* / Q000175:diagnosis; D052279:Creatine Kinase, MB Form / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007527:Isoenzymes / Q000097:blood; D007770:L-Lactate Dehydrogenase / Q000097:blood*; D013058:Mass Spectrometry; D009203:Myocardial Infarction / Q000097:blood* / Q000175:diagnosis; D009211:Myoglobin / Q000097:blood*; D019210:Troponin I / Q000097:blood*
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D007527:Isoenzymes; D009211:Myoglobin; D019210:Troponin I; D007770:L-Lactate Dehydrogenase; C409667:lactate dehydrogenase 1; D052279:Creatine Kinase, MB Form
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