Epitope Localization of Monoclonal Antibodies Used in Human Troponin I Immunoenzymometric Assay

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Abstract
Serum troponin I isoforms have proven to be potent markers of striated muscle injury. They reach the bloodstream soon after their liberation from the damaged muscular cell and can be detected by the use of selected antibodies. Among monoclonal antibodies (MAbs) originally produced against cardiac troponin I (cTnl), two MAbs able to cross react with the skeletal isoform of troponin I (sTnI) were selected and used to develop a one-step immunoenzymometric assay which allows the quantification of both cardiac and skeletal isoforms (scTnl IEMA). The present report describes the main characteristics of this assay. By using multiple peptide synthesis methods, the localization of the epitopes recognized by the two MAbs on sTnI were determined. The capture and tracer MAbs of the scTnl IEMA were shown to recognize epitopes located within positions 121–127 and 160-–167 in the sTnI sequence, respectively. The results of this epitopic analysis are discussed in light of the cross reaction of these two MAbs with cTnl.

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