ESTABLISHMENT OF HYBRIDOMA CELL LINES SECRETING ANTI-TERBUTALINE MONOCLONAL ANTIBODY AND PREPARATION,IDENTIFICATION OF ITS IMMUNOLOGICAL PROPERTIES

Artificial antigen BSA-TBL(terbutaline) and OVA-TBL were synthesized using butane-1,4-diol diglycidyl ether and EDC by linking carrier proteins BSA and OVA to TBL.The titers of polyclonal antibody were detected by indirect ELISA and blocking ELISA after three times immunization by BSA-TBL.The high titer,sensitivity and specificity mouse was selected for cell fusing.The hybridoma lines that secrete TBL mAb were established with using monoclonal antibody hybridoma technology and the immunological characteristics such as titer,sensitivity and specificity of the mAb were characterized.The results showed that six BALB/c mice indirect ELISA titer against TBL were above 1×10-4 and three hybridoma cell lines of 4C08-G5 and 3H3-A02 were screened for specificity to TBL,the indirect ELISA titer of the mAb were 1∶800 and 1∶1600 in supernatant,1∶2.56×105and 1∶1.02×106 in ascites,the mAb of 4C08-G5 showed good sensitivity with an IC50 of 5.25 ng/ml to TBL and had less than 30% cross-reactivity to other compounds.The results showed TBL mAb has been generated and made it possible to establish immunoassay of TBL residues.