Title: Development of a Candidate Secondary Reference Procedure (Immunoassay Based Measurement Procedure of Higher Metrological Order) for Cardiac Troponin I: I. Antibody Characterization and Preliminary Validation
Authors: NOBLE James E.BUNK DavidCHRISTENSON RobertCOLE KennethHE Hua-JunKATRUKHA AlexeiPANTEGHINI M.PORTER Robert A.SCHIMMEL HeinzTATE JillWANG Lili
Citation: CLINICAL CHEMISTRY AND LABORATORY MEDICINE vol. 48 no. 11 p. 1603-1610
Publisher: WALTER DE GRUYTER & CO
Publication Year: 2010
JRC Publication N°: JRC59173
ISSN: 1434-6621
URI: http://publications.jrc.ec.europa.eu/repository/handle/JRC59173
DOI: 10.1515/CCLM.2010.316
Type: Articles in Journals
Abstract: In this study, the first steps in the development of a secondary reference measurement procedure (RMP) "higher metrological order measurement procedure" to support the cardiac troponin I (cTnI) standardization initiative is described. The RMP should be used to assign values to serumbased secondary reference materials (RMs) without analytical artifacts causing bias. A multiplexed bead-based assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were used to identify the optimum monoclonal antibody pair (clones 560 and 19C7) for the RMP. Using these antibodies, an ELISA-based procedure was developed to accurately measure the main cTnI forms present in blood. The proposed RMP appears to show no bias when tested on samples containing various troponin complexes, phosphorylated and dephosphorylated forms, and heparin. The candidate assay displayed suitable linearity and sensitivity (limit of detection, 0.052 mg/L) for the measurement of the proposed cTnI secondary RMs. Preliminary comparison data on patient samples with a commercial cTnI assay are also provided to support the suitability of RMP for value assignment to RMs. Full validation and final assessment of the RMP will be performed through transferability and intercomparison studies.
JRC Institute:Institute for Reference Materials and Measurements

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