Affinity purification and enzymatic cleavage of inter-alpha inhibitor proteins using antibody and elastase immobilized on CIM monolithic disks

摘要: Epoxy-activated monolithic CIM disks seem to be excellent supports for immobilization of protein ligands. The potential use enzymes, immobilized on rapid preparative cleavage proteins in solution was investigated. Digestion complex plasma demonstrated by using inter-alpha inhibitors with elastase, epoxy-activated disks. Recently, a monoclonal antibody against human inhibitor (MAb 69.31) developed. MAb 69.31 blocks the inhibitory activity serine proteases. These results suggest that epitope defined this is located within or proximal active site molecule. This antibody, disk, used very isolation proteins. isolated enzymatic digestion and products, especially from light chain elucidate precisely target sequence N-terminal amino acid sequencing. Bovine pancreatic elastase disk cleaves into small fragments which are still reactive 69.31. One these proteolytic partially sequenced. It could shown at beginning two proteinase domains (bikunin). Elastase offers simple method protease fragments. enzyme stable after repeated runs. A partial complete can achieved varying flow rate.