Human cardiac troponin I (cTnI) is one of the most specific biomarkers for detection of acute myocardial infarction (AMI). To formulate immunoassay kits for rapid immunodiagnosis of AMI, monoclonal antibodies with high affinity and specificity were generated against cTnI and subsequently tested through a series of experiments. C57BL/6 mice were immunized with cTnI as the immunogen and cell fusions with myeloma cells of BALB/c origin were performed to generate hybridomas. The supernatants of the hybridoma cell culture were routinely screened for antibody secretions against intact cTnI and synthetic peptides from the N-terminal half of cTnI (amino acid residues N1-30, N24-40, N59-79, and N80-95). Monoclonal antibodies specific to different epitope regions were then determined and selected, according to their respective affinity and specificity, for formulation of enzyme immunoassay kits. The results of this study found that most of the selected antibodies revealed comparable binding affinity to cTnI and to the corresponding synthetic peptides. Optimal sandwich enzyme immunoassays with high sensitivity could be achieved through proper combinations of the epitope-distinct monoclonal antibodies in different capture-detection pairs; signal enhancements were frequently observed when a mixture of epitope-distinct anti-cTnI monoclonal antibodies was used for coating. This indicates that a combination of epitope-distinct anti-cTnI monoclonal antibodies recognizing the N-terminal half of cTnI yield reliable detection and greater sensitivity for cTnI in AMI patients.

Monoclonal antibodies (MAbs) were generated against subtypes (ad/ad/rw) of the human hepatitis B viral surface antigen (HBsAg). Among dozens of antibodies that were generated, the majority was shown to commonly react with various ad/ay subtypes of the S protein. Epitope(s) of these antibodies were grouped by various immunoassay methods, and at least four distinct epitope regions were identified. Some of these antibodies were selected to formulate sandwich enzyme immunoassays for quantitative determinations of HBsAg in reconstituted specimens. Epitope-defined monoclonal antibodies with high affinity and specificity might be suitable for formulations as vaccines (containing a mixture of humanized monoclonal antibodies) for passive immunization in humans for immunoprophylaxis of HBV infection.