PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST 11S STORAGE PROTEIN FROM PEA-SEEDS

Antibodies can be used as probes to investigate the structure of 11S s torage proteins, their subunit composition and structural modification s induced by technological treatments. Monoclonal antibodies have been raised against Pisum sativum legumin (11S storage protein). Their bin ding characteristics were examined by direct, sandwich and competitive ELISA and by immunoblotting against legumin and 11S type storage prot eins from other species. One of the MAbs. reacting with all 11S protei ns tested, recognizes a discontinuous epitope accessible on the surfac e of the native hexameric protein but destroyed in dissociated legumin . Two antibodies recognize sequential epitopes belonging to a region o f the acidic polypeptides present on the surface of the native legumin . These two MAbs cross-react only with pea and bean 11S proteins. Two other MAbs are specific for sequential epitopes buried in the native p rotein, localized, respectively, on acidic and basic polypeptides. The MAb reacting with the acidic polypeptide exhibits very specific bindi ng for pea legumin. By contrast, the MAb specific to the basic polypep tide cross reacts with 11S proteins studied. This work shows the poten tial of this approach for the characterization of the conformation of the 11S proteins and the investigation of structural modifications.