MONOLITH-BASED PSEUDO-BIOAFFINITY PURIFICATION METHODS FOR FACTOR VIII AND APPLICATIONS THEREOF

The present disclosure relates to purification of Factor VIII protein and/or its fragments from various sources by employing monolith based pseudobioaffinity purification methods. In particular, L-histidine over CIM monolith [Histidine Ligand Affinity Chromatography (HLAC)] is used for the purification of wild-type factor VIII from plasma cryoprecipitate, recombinant B-domain deleted factor VIII (rBDD-FVIII) expressed in various host systems and recombinant factor VIII light chain expressed in Pichia pastoris. Further, immobilized metal over CIM monolith [Immobilized metal-ion affinity chromatography (IMAC)] is employed for the purification of wild-type factor VIII, rBDD-FVIII expressed in various host systems and recombinant factor VIII heavy chain expressed in Pichia pastoris. The purification efficiency showcased by the purification methods of the present disclosure is far superior when compared to the presently available methods for the purification of factor VIII which lead to significantly improved results in therapeutic applications involving factor VIII.