Development of an LC-MRM-MS-Based Candidate Reference Measurement Procedure for Standardization of Serum Apolipoprotein (a) Tests

  • L Renee Ruhaak
    Department of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center , Leiden , The Netherlands
  • Fred P H T M Romijn
    Department of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center , Leiden , The Netherlands
  • Ilijana Begcevic Brkovic
    Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig , Leipzig , Germany
  • Zsusanna Kuklenyik
    Division of Laboratory Sciences, Centers for Disease Control and Prevention , Atlanta, GA , USA
  • Julia Dittrich
    Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig , Leipzig , Germany
  • Uta Ceglarek
    Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig , Leipzig , Germany
  • Andrew N Hoofnagle
    Department of Laboratory Medicine and Pathology, University of Washington , Seattle, WA , USA
  • Harald Althaus
    Siemens Healthcare Diagnostics Products GmbH , Marburg , Germany
  • Eduardo Angles-Cano
    French Institute of Health and Medical Research (Inserm), Université Paris Descartes , Paris , France
  • Stefan Coassin
    Department of Genetics and Pharmacology, Institute of Genetic Epidemiology, Medical University of Innsbruck , Innsbruck , Austria
  • Vincent Delatour
    Laboratoire National de Métrologie et d’Essais , Paris , France
  • Liesbet Deprez
    European Commission, Joint Research Centre , Geel , Belgium
  • Ioannis Dikaios
    European Commission, Joint Research Centre , Geel , Belgium
  • Gerhard M Kostner
    Division of Molecular Biology and Biochemistry, Gottfried Schatz Research Center (for Cell Signaling, Metabolism and Aging), Medical University of Graz , Graz , Austria
  • Florian Kronenberg
    Department of Genetics and Pharmacology, Institute of Genetic Epidemiology, Medical University of Innsbruck , Innsbruck , Austria
  • Alicia Lyle
    Division of Laboratory Sciences, Centers for Disease Control and Prevention , Atlanta, GA , USA
  • Urban Prinzing
    Roche Diagnostics GmbH , Penzberg , Germany
  • Hubert W Vesper
    Division of Laboratory Sciences, Centers for Disease Control and Prevention , Atlanta, GA , USA
  • Christa M Cobbaert
    Department of Clinical Chemistry and Laboratory Medicine, Leiden University Medical Center , Leiden , The Netherlands

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<jats:title>Abstract</jats:title><jats:sec><jats:title>Background</jats:title><jats:p>Medical results generated by European CE Marking for In Vitro Diagnostic or in-house tests should be traceable to higher order reference measurement systems (RMS), such as International Federation of Clinical Chemistry and Laboratory Medicine (IFCC)-endorsed reference measurement procedures (RMPs) and reference materials. Currently, serum apolipoprotein (a) [apo(a)] is recognized as a novel risk factor for cardiovascular risk assessment and patient management. The former RMS for serum apo(a) is no longer available; consequently, an International System of Units (SI)-traceable, ideally multiplexed, and sustainable RMS for apo(a) is needed.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>A mass spectrometry (MS)-based candidate RMP (cRMP) for apo(a) was developed using quantitative bottom-up proteomics targeting 3 proteotypic peptides. The method was provisionally validated according to ISO 15193 using a single human serum based calibrator traceable to the former WHO-IFCC RMS.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>The quantitation of serum apo(a) was by design independent of its size polymorphism, was linear from 3.8 to 456 nmol/L, and had a lower limit of quantitation for apo(a) of 3.8 nmol/L using peptide LFLEPTQADIALLK. Interpeptide agreement showed Pearson Rs of 0.987 and 0.984 for peptides GISSTVTGR and TPENYPNAGLTR, and method comparison indicated good correspondence (slopes 0.977, 1.033, and 1.085 for LFLEPTQADIALLK, GISSTVTGR, and TPENYPNAGLTR). Average within-laboratory imprecision of the cRMP was 8.9%, 11.9%, and 12.8% for the 3 peptides.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>A robust, antibody-independent, MS-based cRMP was developed as higher order RMP and an essential part of the apo(a) traceability chain and future RMS. The cRMP fulfils predefined analytical performance specifications, making it a promising RMP candidate in an SI-traceable MS-based RMS for apo(a).</jats:p></jats:sec>

収録刊行物

  • Clinical Chemistry

    Clinical Chemistry 69 (3), 251-261, 2023-01-16

    Oxford University Press (OUP)

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