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

Artificial antigen BSA-CIM and OVA-CIM were synthesized using diazotization by linking carrier proteins BSA and OVA to CIM.The titer of polyclonal antibody(pAb) was detected by indirect ELISA and blocking ELISA after three times immunization by BSA-CIM.The high titer,sensitivity and specificity mouse was selected for cell fusing.The hybridoma lines that secrete CIM mAb were established using monoclonal antibody hybridoma technology and the immunological characteristics such as titer,sensitivity and specificity of mAb were characterized.The results showed that six BALB/c mice indirect ELISA titer against CIM were above 1 × 10-3 and three hybridoma cell lines of 3B11-H4,2A11-G11 and 4F5-F11were screened for specificity to CIM,the indirect ELISA titer of the mAb were 1 ∶ 800,1 ∶ 1600and 1∶ 1600 in supernatant,1 ∶ 2.56 × 106,1 ∶ 1.02 × 107 and 1 ∶ 2.56 × 107 in ascites,the mAb of 3B11-H4 showed good sensitivity with an IC50 of 0.40ng/ml to CIM and had less than 0.2% cross-reactivity to other compounds.The results showed the high-titer,sensitivity and specificity CIM mAb has been generated and made it possible to establish immunoassay of CIM residues.