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| Preparation and Characterization of Monoclonal Antibody against Sheep Skeletal Muscle Troponin T |
| LIU Jingjing, WANG Zhenhua, LI Suhong, ZHANG Jing, WU Meng, DU Shunfeng, ZHANG Yan, LI Chunsheng |
| 1.Biology Institute of Hebei Academy of Sciences, Shijiazhuang 050081, China; 2.Hebei Kexing Pharmaceutical Co. Ltd., Shijiazhuang 050200, China; 3.Baoding Animal Product Quality Inspection and Monitoring Center, Baoding 071027, China; 4.Qinhuangdao Agricultural Products Quality and Safety Supervision and Inspection Center, Qinhuangdao 066001, China; 5.Hebei Food Inspection and Research Institute, Shijiazhuang 050227, China |
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Abstract Monoclonal antibody against sheep skeletal muscle troponin T (sTnT) was prepared and its properties were evaluated. For this aim, sTnT was prepared by extraction in boiling NaCl solution and purified by sodium dodecyl sulfate polyacrylamide gel electropheresis. Hybridoma cell lines with stable secretion of sTnT were obtained after fusion. The titer, subtype and specificity of monoclonal antibodies were analyzed by enzyme linked immunosorbent assay. Three stable monoclonal antibodies (2E11, 3D3 and 4C2) were obtained after cloning. Among these, 4C2 had the best specificity with a titer of 1:5.12 × 105. The subtype of 4C2 was IgG1 and its affinity constant was 1.1 × 109 L/mol. Western blot results showed that the monoclonal antibody reacted with bovine and sheep skeletal muscle extracts but not chicken and duck skeletal muscle extracts. Accordingly, 4C2 could allow discrimination of ruminant from poultry skeletal muscle.
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2020, Vol. 34 
