DEVELOPMENT OF A FREEZE-DRIED ALBUMIN-FREE FORMULATION OF RECOMBINANTFACTOR-VIII SQ

Purpose. To develop a stable freeze-dried formulation of recombinant f actor Vm-SQ (r-VIII Sa) without the addition of albumin, Methods, Diff erent formulations were evaluated for their protective effect during s terile filtration, freeze-thawing, freeze-drying, reconstitution and l ong term storage. Factor VIII activity (VIII:C), visual inspection, cl arity, solubility, moisture content and soluble aggregates and/or frag ments were assayed,Results, A combination of non-crystallising excipie nts (L-histidine and sucrose), a non-ionic surfactant (polysorbate XO) and a crystalline bulking agent (sodium chloride) was found to preser ve the factor VIII activity during formulation, freeze-drying and stor age, Calcium chloride was included to prevent dissociation of the heav y and light chains of r-VIII SQ, Sodium chloride was chosen as the pri mary bulking agent since the concentration of sodium chloride necessar y for dissolution of r-VIII SQ in the buffer will inhibit the crystall ization of many potential cake formers, It was found that L-histidine, besides functioning as a buffer, also protected r-VIII SQ during free ze-drying and storage. A pH close to 7 was found to be optimal. Some p otential macromolecular stabilisers, PEG 4000, Haes(R)-steril and Haem accel(R), were evaluated but they did not improve the recovery of VIII :C. The freeze-dried formulation was stable for at least two years at 7 degrees C and for at least one year at 25 degrees C. The reconstitut ed solution was stable for at least 100 hours at 25 degrees C. Conclus ions. The albumin-free formulation resulted in consistently high recov ery of VIII:C, very law aggregate formation and good storage stability , The stability of the reconstituted solution makes the formulation su itable for continuous administration via infusion pump, The formulatio n strategy described here may also be useful for other proteins which require a high ionic strength.