Isolation of intact proteins from acid-degradable polyacrylamide gel

Elucidating native structure-function relationships of proteins identified using PAGE has been impeded by limitations in the isolation of intact proteins from the gel. By hydrolyzing polyacrylamide gel band under mildly acidic conditions rather than digesting entrapped proteins approximately 70% of a large native protein, mouse IgG1 (molecular weight 150 kDa), was isolated. Further analysis indicated that the isolated antibodies had preserved specific binding capability to target antigens as well as intact molecular weights. This new technology may contribute to functional proteomic studies through the isolation of proteins in their native state after PAGE, and other technologies requiring simultaneous separation and isolation of other macromolecules and complexes.