Preparation and immunological trait of monoclonal antibody for sulfadiazine.

【Objective】The monoclonal antibodis against Sulfadiazine were prepared and their immunological characteristics were identified.【Method】Diazotization was used to conjugate SD to carrier protein BSA and artificial antigen was obtained.The coating antigen OVA-SD was obtained in the same way,ultraviolet(UV)and SDS-PAGE were used to identify SD artificial antigen.BALB/C mice were immunized with BSA-SD,the titre of polyclonal antibody(pAb) was detected by indirect ELISA and blocking ELISA.The hybridoma lines that secrete SD mAb were established with monoclonal antibody hybridoma technology.The immunological characteristics such as titer,sensitivity and specificity of the mAb were studied.【Result】The results showed that SD artificial antigen was synthesized successfully and its conjugation ratio of SD to BSA was about 10.3∶1,three hybridoma cell lines of 6B5-E4、6B5-E9、3A5-B4 and 2H8-G5 were screened for specificity to SD,the indirect ELISA titer of the mAb were 1∶810,1∶256,1∶512,1∶512 and in supernatant,1∶(8.6×106),1∶(2.5×106),1∶(5×106),1∶(5×106) in ascites,the mAb of 6B5-E4 showed good sensitivity with an IC50 of 0.54 μg/L to SD,2.7% cross-reactivity to Sulfamonomethoxine and little or no cress-reactivity to other compounds.【Conclusion】SD mAb of high-titer,sensitivity and specificity has been generated and made it possible to establish immunoassay of SD residues.