Cardiac troponin I (cTnI) is the biomarker of choice
and considered
a gold standard for the diagnosis of acute myocardial infarction.
However, the quantitative results of cTnI assay kits from different
manufacturers are not comparable. Based on the H/D exchange mass spectrometry
(HDX-MS) workflow, we developed an in-vitro diagnostic reagent antibody
evaluation strategy to analyze the interactions of epitopes and antibody
cocktails(R195, F12, S13) and (D1, D2, pAb2). The HDX results
indicate that the quantitative result bias of the different reagents
originates from the ability of antibodies to recognize various cTnI
complex forms, such as free cTnI, hydrolyzed cTnI, and cTnI combined
with cTnT or TnC as binary or ternary complexes (cTnIC, cTnTIC), in
blood based on different epitopes. The data obtained from the peptide
HDX of interest after treatment with various antibody cocktails clearly
indicated epitope specificity. The consistency of quantitative results
can be improved by a thorough investigation into the epitopes recognized
by the antibodies of various diagnostic kits, which will lead to the
standardization of cTnI diagnosis.
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