Affinity chromatography of human blood coagulation factor VIII on monoliths with peptides from a combinatorial library

FVIII is a very complex molecule of great therapeutic significance. It is p urified by a sequence of chromatographic steps including immunoaffinity chr omatography. A peptide affinity chromatography method has been developed us ing peptides derived from a combinatorial library. Spot technology using ce llulose sheets has been applied for this purpose. The dual positional scann ing strategy was used for identification of the amino acids in random posit ions. Approximately 5000 possible candidates found in the first screening r ound were reduced to a panel of 36, Six candidates have been selected empir ically, Five peptides seem to be directed against the light chain of FVIII, one peptide seems to be directed against the heavy chain. The peptides have been immobilized on conventional beaded material and CIM polymethacrylate monoliths. Much better performance with respect to capacit y and selectivity has been observed with the monolithic material. Exposure of the ligand and its ensuing accessibility are responsible for these prope rties.