Hemophilia A is an inherited bleeding disorder that stems from lack of coagulation factor VIII (F.VIII). Crude preparations of plasma-derived F.VIII used in treatment of hemophilia are in short supply worldwide and carry the risk of viral disease transmission. Although ultrapurified F.VIII can be prepared using immunoaffinity column chromatography, conventional affinity separation technology is poorly suited to large-scale production of purified F.VIII.The objective of this research is to develop a membrane-based affinity process for the purification of F.VIII directly from blood plasma. The membrane affinity processes will offer high specificity and yield, and yet be amenable to industrial operations.In Phase I, the technical feasibility of membrane immunoaffinity purification of F. VIII from plasma was established.In Phase II, the F.VIII purification process will be optimized and scaled up for production of materials for preclinical studies. F.VIII thus purified will be extensively characterized and tested for safety in animals. The purpose of this project is to establish the technical basis for membrane-mediated affinity purification of F.VIII, to develop a general approach to industrial affinity separations, and to improve the quality of life of individuals undergoing F.VIII replacement therapy.Awardee's statement of the potential commercial applications ofthe esearch: This research may lead to development of(1) a safer, more readily available, and less expensive F.VIII product that could address the needs of individuals undergoing replacement therapy, and(2) a general approach to industrial purification of therapeutic products derived from either blood plasma, fermentation media, or cell culture broths.National Heart, Lung, and Blood Institute (NHLBI)