Anti-idiotypic monoclonal Ig specific for an anti-laminin Ig heavy chain transgene variable region.
Techniques currently available to obtain anti-idiotypic reagents reactive with a single chain of a lymphocyte antigen receptor rely on immunization with intact soluble or cell-bound Ig or T-cell receptors. Ready recovery of single-chain-specific monoclonal antibodies (MAbs) depends on the presence of an immunodominant epitope on the desired chain and chance recovery of the responding clone. Here we present a method to maximize recovery of an Ig heavy-chain-specific anti-idiotypic Ig, using sequential immunization with MAbs expressing the H chain V region in combination with different H chain isotypes and with different light chains. The latter was produced by in vitro transfection of an H-chain-loss variant myeloma cell line with a transgene construct expressing the Ig H chain V region of interest. Sequential immunization may be a useful strategy to enhance selection of anti-Id reagents reactive with single chain-specific epitopes.