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| Preparation and Characterization of Monoclonal Antibody against Duck Skeletal Muscle Troponin I |
| LIU Jingjing, ZHANG Jing, SUN Jinchong, WU Meng, DU Shunfeng, LI Chunsheng |
| 1.Biology Institute of Hebei Academy of Sciences, Shijiazhuang 050081, China; 2.Qinhuangdao Agricultural Products Quality and Safety Supervision and Inspection Center, Qinhuangdao 066001, China |
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Abstract A monoclonal antibody (mAb) against duck skeletal muscle troponin I (sTnI) was prepared and its characteristics were evaluated. sTnI was analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE). The protein band was excised from the gel and used to immunize mice. Hybridoma cell strains stably secreting mAb were obtained by fusion. The titer, subtype and speci?city of monoclonal antibodies were analyzed by enzyme linked immunosorbent assay. The SDS-PAGE showed 2 dark bands. The 24 kDa protein was selected for immunization. Two cell lines that secreted monoclonal antibody against sTnI were obtained after screening, with 3E7 being the one with higher speci?city. It belonged to the IgG1 subtype, its titer was 1:1.0 × 105 and its af?nity constant was 5.9 × 105 L/mol. Western blot results showed that 3E7 reacted with duck skeletal muscle extracts from duck, but not bovine, sheep, chicken and ?sh skeletal muscle extracts.
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2020, Vol. 34 
