Self-immobilizing recombinant antibody fragments for immunoaffinity chromatography: generic, parallel and scaleable protein purification. Protein Expt

We present the directed immobilization of recombiof great value in the rapid purification of proteins to high degrees of purity. The antibody could either be nant antibody fragments as ligands for general immunoaffinity chromatography methods. It is based on fusion proteins of scFv fragments with several chitinbinding domains which can be immobilized directly from a crude bacterial lysate on inexpensive chitin beads for the purification of proteins without any gradient or detector. It has been used with a positive pressure manifold, allowing the parallel processing of 24 differ-ent samples on a milligram scale, as convenient as plasmid isolation. The method is demonstrated with several anti-protein antibodies. In addition, methods are presented of using an anti-His tag antibody either alone o