Production of anti‐idiotypic monoclonal antibodies that mimic the phytotoxin dothistromin

Abstract

Dothistromin (DOTH) is a toxin secreted by Dothistroma pini and is present in needle lesions resulting from infection by this fungus. A monoclonal antibody (MAb 10C12) of high affinity for DOTH was conjugated to bovine serum albumin (BSA) and used to immunize mice in order to prepare monoclonal anti‐idiotypic antibody mimics of dothistromin. One anti‐idiotypic antibody, 12C9G8, was equivalent to DOTH on a molar basis in inhibition of the binding of an anti‐DOTH MAb to DOTH‐protein conjugates. Furthermore, in a Western blot analysis, 12C9G8 visualized the same protein profile in embryos as was observed in an approach which utilized an anti‐DOTH MAb together with DOTH‐mouse serum albumin (MSA) conjugate to identify potential DOTH binding sites. In a parallel immunogold electron microscopy experiment, protein‐containing vesicles were labelled using both approaches.

Keywords:

• Dothistromin
• anti‐idiotypic mimic
• monoclonal antibodies
• ELISA