Preparation and Immunological Characterization of Monoclonal Antibody against 2,4-D

The aim of the study is to prepare high sensitivity and specificity monoclonal antibody(mAb) against 2,4-D and identify its immunological characteristics.The 2,4-D was conjugated to carrier protein BSA and OVA by EDC to obtain the immunogen and reactionogen,and SDS-PAGE was used to identify these artificial antigens.Balb/c mice were immunized with the immunogen.The high titer and sensitivity mouse was selected for cell fusing by indirect ELISA and blocking ELISA after four times immunization;the hybridoma lines that secrete 2,4-D mAb were established by using monoclonal antibody hybridoma technology and the immunological characteristics such as titer,sensitivity,subclass and specificity of the mAb were characterized.Three hybridoma cell lines,1F11,2A12 and 2G12,were screened for specificity to 2,4-D after cell fusing.All the mAb against 2,4-D were type IgG1,the titer of the mAb were 1∶2 560-1∶5 120 in supernatant,the mAb of them showed good sensitivity with IC50 of 0.801 μg/L,1.332 μg/L,1.564 μg/L to 2,4-D,and the affinity constant(Ka) were 2.174×1011 L/mg,3.21×1010 and 4.36×1010 L/mg,respectively.The rates of cross reaction of the mAb with other compounds were less than 0.03%.The research has successfully obtained the high sensitivity,high affinity and high specificity of 2,4-D mAb,it has established solid foundation for 2,4-D immunological rapid detection method.