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
Articolo
Data di Pubblicazione:
2010
Citazione:
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 / J. E. Noble, D. M. Bunk, R. H.
Christenson, K. D. Cole, H. J. He,
A. G. Katrukha, M. Panteghini, R. A.
Porter1, H. Schimmel, J. R. Tate, L. Wang on behalf of the IFCC Working Group on Standardization of Troponin ( WG-TNI). - In: CLINICAL CHEMISTRY AND LABORATORY MEDICINE. - ISSN 1434-6621. - 48:11(2010), pp. 1603-1610. [10.1515/CCLM.2010.316]
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
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
Antibody specificity; Antigen-antibody reactions; Candidate reference procedure; ELISA; Reference standards; Troponin I
Elenco autori:
J.E. Noble, D.M. Bunk, R.H. Christenson, K.D. Cole, H.J. He, A.G. Katrukha, M. Panteghini, R.A. Porter1, H. Schimmel, J.R. Tate, L. Wang on behalf of the IFCC Working Group on Standardization of Troponin
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