Isolation of alpha-crystallin and its subunits by affinity chromatography on immobilized monoclonal antibodies

Use has been made of the specific interactions between monoclonal antibodies and the alpha A or alpha B subunits of alpha-crystallin to devise methods for the purification of the intact protein or its subunits. alpha A and alpha B subunits were separated by affinity chromatography on an immobilized monoclonal antibody specific for alpha A chains, using a pH gradient. Use of an antibody which binds both subunits has enabled the isolation of intact alpha-crystallin aggregates. Gel electrophoresis, conformational probing and size analysis showed that the affinity purified proteins were purer but otherwise indistinguishable from the alpha-crystallins isolated by other methods.