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| Preparation and Characterization of Monoclonal Antibody against Porcine Skeletal Muscle Troponin T |
| ZHANG Jing, LI Yujing, ZHANG Erjing, LI Ziran, LIU Jingjing, LI Chunsheng |
| 1. Institute of Biology, Hebei Academy of Science, Shijiazhuang 050081, China; 2. Hebei Jiaotong Vocational and Technical College, Shijiazhuang 050035, China; 3. College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China |
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Abstract In order to prepare monoclonal antibodies against porcine skeletal muscle troponin T (sTnT), porcine skeletal muscle troponins were extracted by biochemical methods and used to immunize mice. By means of cell fusion, we acquired hybridoma cell strains that can secrete monoclonal antibodies (mAbs) stably. Ascites was induced in vivo to further prepare and characterize monoclonal antibodies against sTnT. Finally, we screened out one hybridoma cell strain that secretes mAb stably and named it 2E8. The monoclonal antibody 2E8 belonged to the IgG2a subtype, whose titer and affinity constant were 1 : 4.1 × 105 and 5.14 × 106 L/mol, respectively. The specificity of 2E8 was evaluated by indirect enzyme-linked immunosorbent assay (iELISA). The cross-reactivity with chicken sTnT and duck sTnT were both 25%. The cross-reactivity with bovine sTnT and goat sTnT were 1.7% and 6.3% respectively. Immunoblotting data showed that the monoclonal antibody 2E8 exhibited a clear band after reaction with the porcine antigen, but only a slight band after reaction with the goat, chicken, or duck antigen. In addition, the monoclonal antibody hardly reacted with the bovine antigen.
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2021, Vol. 35 
