Preparation of Monoclonal Antibodies and Development of an Indirect Competitive ELISA for Fumonisin B_1 detection

To establish an indirect competitive ELISA for Fumonisin B1 detection,conjugations of KLH-FB1 and OVA-FB1 were synthetized using glutaraldhyde method and they were employed as the coating antigen and immunogen,respectively.8-week-old Balb/c mouse was immunized by the KLH-FB1 to develop the monoclonal antibody(MAb) using hybridoma technique.After three times of subcloning,two hybridoma cell lines stably secreting specific antibody against FB1 were screened by indirect competitive ELISA.The ascites MAb were purified and characterized by SDS-PAGE.The titre of the MAb reached 1∶16 000.In addition,the immunological sub-type of the MAb was identified as IgG1 and the its light chain belonged to κ type.Aflatoxin B1,Chlorpromazine,streptomycin and Zearalenone showed no cross-activity with the monoclonal antibody.Indirect competitive ELISA(IC-ELISA) was established based on the specific antibody against FB1.The sensitivity of IC-ELISA was 0.44 ng/mL and the liner range was 0.93~73.06 ng/mL.The method was used to detect FB1 in spiked samples and the recoveries were 78.4%~102% in the detection range.