Acridinium Ester Labelling Nd Characterization of Bovine Cardiac Troponin (cTn-I) Specific Monoclonal Antibodies

The use of monoclonal antibody (McAb) based immunoassays to detect serum concentration changes of cardiac specific structural and regulatory proteins, as biochemical markers, is gaining immense interest in diagnostic procedures for cardiomyocyte necrosis. Of the non-isotopic probes, chemiluminescent molecules such as acridinium esters (A.E.) are regarded as being of real value in routine clinical chemistry. In the present study, bovine cardiac troponin-I specific 5F4 McAb was purified by Protein 'A' and Mono-Q ionexchange chromatography and conjugated with A.E. The method of purification hardly interfered with the conjugation procedure. The A.E. conjugated 5F4 retained its immunoreactivity post conjugation and was used to design direct and indirect immunoassays for the detection of cTn-I in vitro samples.