Stability of high concentration rhuMAb VEGF

BIOT 13

Hong Liu, hongliu@gene.com and Mary E. M. Cromwell, cromwell@gene.com. Early Stage Pharmaceutical Development, Genentech, Inc, 1 DNA Way 96A, South San Francisco, CA 94404
The purpose of this study was to develop a high concentration rhuMAb VEGF formulation for subcutaneous (SC) delivery. A stability study was designed with 100mg/ml rhuMAb VEGF, based on rhuMAb VEGF's unique reversible self-association. The extent of self-association depends on pH, protein concentration and ionic strength. Formulations were tested over a narrow pH range to minimize aggregation. Protein stability in these formulations was evaluated at 2-8°C and –20°C storage by Size Exclusion HPLC and Ion Exchange HPLC. Due to concern of potential hydrolysis of sucrose under the lower pH conditions, a boronate affinity HPLC method was utilized to evaluate glycation on stability samples.

The study shows that lower pH can help to minimize the amount of aggregate at a rhuMAb VEGF concentration of 100mg/ml. As expected, sugars helped in freeze/thaw stability and storage at –20°C. There are significant differences in aggregate content between sucrose-containing and trehalose -containing liquid formulations stored at 2-8°C, and the differences are not due to glycation.