Chromatographic separation of proteins using polyol-responsive monoclonal antibodies.

This technique takes advantage of the unique properties of a class of monoclonal antibodies, termed polyol-responsive mAbs. These can be used in immunoaffinity purification. The singular advantage of the Polyol-Responsive mAb System is that the target protein is not denatured during purification, so it is suitable for the purification of labile, multi-subunit protein complexes. These protein complexes bind tightly to the polyol-responsive mAbs, but they are released under very gentle elution conditions.

The mAbs can be attached to beads without inactivating the antibodies. When a mixture of different proteins in normal buffer is combined with polyol-responsive mAb-bound beads, the target protein binds to the mAbs. The other proteins remain in solution. In the batch mode, after filtration, the target-protein-mAb bead combination is resuspended in the appropriate polyol-containing buffer, releasing the target protein. A second filtration yields the pure, target protein with excellent recovery and enzyme activity.

These polyol-responsive mAbs can also be used in a column chromatography.

Detailed descriptions and protocols of the uses of polyol-responsive mAbs are listed in protocols, also look at references.