Extraction, Purification And Identification Of Bovine Myocardial Troponin T And Preparation Of Corresponding Monoclonal Antibodies

Objective: This research is aimed at obtaining mouse-anti-bovine myocardial troponin T (cTnT) monoclonal antibodies through immunizing experimental animals with extracted, purified and identified bovine cTnT.Methods: The extraction and purification of cTnT was practiced through tissue homogenate and protein chromatography procedures. Obtained protein was identified through SDS-PAGE and ELISA. After animal immunization step, the classical cell fusion technique was employed to produce hybridomas.Selected positive hybridoma,which was determined after corresponding supernatant was detected though ELISA with purified cTnT,was isolated ,mono-cloned, expanded and cryopreservated. The titer of ascites collected after injecting hybridomas into the mouse abdominal cavity, was evaluated through indirect ELISA.Results: SDS-PAGE demonstrated the molecular mass of the obtained protein was 37KD, with a highest band proportion of 93.77% in corresponding gel track. Specific combination between obtained protein and cTnT monoclonal antibody was revealed through ELISA. After cell fusion and isolation, five hybridoma strains, namely 1C9, 1F2, 2B8, 3A6,3B10, which can stationarily secrete monoclonal antibodies, have been expanded and cryopreservated.Conclusion: High purity cTnT protein and mouse-anti-bovine cTnT monoclonal antibodies were obtained, which established the fundamental support for further development.