Rapid and sensitive procedure for assigning idiotypic determinants to heavy or light chains: Application to idiotopes associated with the major cross-reactive idiotype of A/J anti-phenylarsonate antibodies

A rapid and sensitive procedure is described for assigning idiotypic determinants to heavy or light polypeptide chains. Heavy and light chains are resolved by electrophoresis in the presence of sodium dodecyl sulfate. The electrophoretically resolved polypeptides are then transferred to nitrocellulose filters. Filters containing bound heavy and light chains are incubated with 125I-labelled anti-idiotypic antibody, and idiotype-anti-idiotype reactivity visualized by autoradiography. This procedure is illustrated with three monoclonal anti-idiotopic antibodies which recognize determinants associated with the major cross-reactive idiotype family of A/J anti-phenylarsonate antibodies. All three anti-idiotopic antibodies are shown to react with electrophoretically resolved idiotype heavy chain, but not with idiotype light chain.