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

Published

November 1, 2010

Author(s)

James E. Noble, David M. Bunk, R. H. Christenson, Kenneth D. Cole, Hua-Jun He, Alexei Katrukha, Mauro Panteghini, Robert Porter, Heinz Schimmel, Jillian Tate, Lili Wang

Abstract

The development of a reference measurement procedure to support the cardiac troponin I (cTnI) standardization initiative is described. The reference measurement procedure will be used to assign reference values to serum-based secondary standards. A multiplexed bead-based assay and SDS-PAGE were used to identify the optimum antibody pair; clones 560 and 19C7 for the reference assay. Described is current progress on the development of the ELISA based procedure that will need to accurately measure all isoforms of cTnI present in serum. The current RMP appears to show no bias when tested with purified samples representing various troponin complexes, phosophorylated states and heparin interference. The candidate assay displays suitable linearity and sensitivity (limit of detection of 52 ng/L) for the measurement of the proposed cTnI serum standards. Limited data with patient samples and comparison with commercial analyzers is provided to support its suitability for reference value assignment. Full validation and assessment of the procedure will be determined through round-robin studies.

Citation

Clinical Chemistry and Laboratory Medicine

Volume

48

Issue

11

Pub Type

Journals

Keywords

Antigen-Antibody Reactions, Antibody Specificity, Candidate reference procedure, ELISA, Reference standards, Troponin I.

Citation

Noble, J. , Bunk, D. , Christenson, R. , Cole, K. , He, H. , Katrukha, A. , Panteghini, M. , Porter, R. , Schimmel, H. , Tate, J. and Wang, L. (2010), 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, Clinical Chemistry and Laboratory Medicine (Accessed October 17, 2025)