Preventing Aggregation of Recombinant Interferon beta-1b in Solution by Additives: Approach to an Albumin-Free Formulation

Abstract

Purpose: Aggregation suppressing additives have been used to stabilize proteins duringmanufacturing and storage. Interferonβ-1b is prone to aggregation because of being nonglycosylated.Aggregation behavior of albumin-free formulations of recombinant IFNβ-1bwas explored using additives such as n-dodecyl-β-D-maltoside, Tween 20, arginine,glycine, trehalose and sucrose at different pH.Methods: Fractional factorial design was applied to select major factors affectingaggregation in solutions. Box-Behnken technique was used to optimize the bestconcentration of additives and protein.Results: Quadratic model was the best fitted model for particle size, OD350 andOD280/OD260. The optimal conditions of 0.2% n-Dodecyl-β-D-maltoside, 70 mMarginine, 189 mM trehalose and protein concentration of 0.50 mg/ml at pH 4 were achieved.A potency value of 91% ± 5% was obtained for the optimized formulation.Conclusion: This study shows that the combination of n-Dodecyl-β-D-maltoside, arginineand trehalose would demonstrate a significant stabilizing and anti-aggregating effect on theliquid formulation of interferonβ-1b. It can not only reduce the manufacturing costs but willalso ease patient compliance.

Keywords: n-Dodecyl-β-D-maltoside, Optimization, HSA-free formulation, Aggregation, Box-Behnken experimental design