Preparation of monoclonal antibody against Shiga-like toxin type II variant B.

To establish a new detection method for Shiga-like toxin type Ⅱ variant(SLT-Ⅱ e),the SLT-Ⅱ e B gene was amplified by PCR and aequence compared with reference strains.The gene was subcloned into pET-30a vector for expression and the recombinant product of SLT-Ⅱe B was used to immunize the BALB/c mice.Six hybridoma cell lines secreting monoclonal antibodies(McAbs) against SLT-Ⅱe B were established,The McAbs belonged to IgM,IgA,IgG2b and IgG2a subtype with κ chain.Blocking ELISA confirmed that all McAbs were able to reacted with native SLT-Ⅱ e.Addition ELISA confirmed that McAbs 3G7 and 4F3 recognized different epitopes.