The purpose of this study was to examine the effects of adsorption of recombinant human interferon-gamma (rhIFN-gamma) on ice surfaces and subsequent drying during processing by spray-lyophilization and lyophilization. Ice/liquid interfacial areas were manipulated by the freezing method as well as by the addition of an annealing step during lyophilization; that is, rhIFN-gamma adsorption was modified by the addition of nonionic surfactants. rhIFN-gamma was lyophilized or spray-lyophilized at a concentration of 1 mg/mL in 5% sucrose, 5% hydroxyethyl starch (HES) +/- 0.03% polysorbate 20 in 140 mM KCl, and 10 mM potassium phosphate, pH 7.5. After the samples were frozen, half were annealed on the lyophilizer shelf. Recovery of soluble protein was measured at intermediate points during processing. On drying, the secondary structure of rhIFN-gamma was determined by second-derivative infrared (IR) spectroscopy, specific surface areas (SSAs) were measured, scanning electron micrographs (SEM) were taken, and dissolution times were recorded. Adsorption of rhIFN-gamma to ice/liquid interfaces alone was not responsible for aggregation. Rather, drying was necessary to cause aggregation in lyophilized sucrose formulations. Addition of an annealing step to the lyophilization cycle resulted in more native-like secondary protein structure in the dried solid, eliminated cracking of the dried cakes, and suppressed both the formation of air/liquid interfaces and rhIFN-gamma aggregation on reconstitution.
Copyright 2003 Wiley-Liss, Inc. and the American pharmaceutical Association J Pharm Sci 92:715-729, 2003