Abstract：BALB/c mice were immunized intra-peritoneally with the fusion protein Eac-Stx1/2B which had been purified by SDS-PAGE and electroelution,and the whole bacteria of another constructed recombinant strain Δler/str(pBR322::stxl/2B::eae).Spleen cells collected from the immunized mice were fused with mouse myeloma cells Sp2/0.Three hybridoma cell lines specific to E.coli O157 were obtained after screening with indirect ELISA and subcloning.They were designated as 2H9,3B4 and 4H7 respectively.Identification of the subclass indicated that the monoclonal antibody secreted by 2H9 cell line belonged to IgG2a.while those secreted by 3B4 and 4H7 belonged to IgM.As demonstrated by indirect ELISA,the titers of the purified ascetic fluids of 2H9,3B4 and 4H7 were 1∶1.28×106,1∶3.2×104 and 1∶6.4×104,and the affinity constants of these antibodies were 2.9×106,1.4× 106 and 2.6 x 105 respectively.It is evident that these monoclonal antibodies all show high degree of specificity in which 2H9 antibody has the highest titer and affinity constant.
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