Abstract: 【Objective】 The aim of the study is to generate high sensitivity and high specificity monoclonal antibody against kasugamycin and identify its immunological characteristics.【Method】KSM was coupled with the carrier protein BSA and OVA by using EDC method and the immunogen KSM-BSA and coating antigen KSM-OVA were synthesized respectively, and SDS-PAGE was used to identify KSM-BSA. Balb/c mouse was immunized by using KSM-BSA. Immunization interval was 4 weeks. The titer of polyclonal antibody was detected by indirect ELISA and blocking ELISA after four times immunization. The high titer and high sensitivity mouse was selected for cell fusion. High sensitivity and high specificity monoclonal antibody was prepared after several times subcloning,and the immunological characteristics were characterized. The preparation of ascites was carried out using in vivo induction method. 【Result】 SDS-PAGE results showed that KSM-BSA artificial antigen was synthesized successfully.Indirect ELISA showed a high titer above 1:104 of the antiserum of all the six Balb/c mouse. The sensibility of antiserum of No.3 mice was the best, which IC50 was 73.9ng•mL-1. Hybridoma cell lines of 1G7 and 2C8 were screened after cell fusion, all the isotypes of the mAb were IgG1.Titers of the mAb were 1:5.12×103 and 1:1.28×103 in supernatant,1:5.12×105 and 1:2.56×105 in ascites. The IC50 of KSMmAb was 8.902 ng•mL-1 to kasugamycin. The rate of cross reaction of the mAb with other compounds was less than 0.03%.【Conclusion】KSM artificial antigens were synthesized successfully and the monoclonal antibody with high sensitivity and specificity was obtained. This study has laid a foundation for a KSM immunological fast detection method.

Key words: kasugamycin , artificial antigen , Monoclonal antibody