Formulating appropriate storage conditions for biopharmaceutical proteins is essential in ensuring the potency, efficacy and stability of these substances over their shelf-life. A typical purification scheme for monoclonal antibodies includes Protein A chromatography with an output of concentrated antibody in a low pH solution. Previously, a purified model antibody was formulated in an acetate buffer that caused aggregation, precipitation, and antibody loss over time, which was exacerbated by freeze/thaw cycles. Other common buffer systems like Tris, citrate and histidine were similarly unacceptable. In order to improve stability of this challenging murine IgG3, we used a 23 full factorial design of experiments (DoE) approach to analyze different formulations of 3 buffer systems including acetate, arginine, and histidine. The formulations were assessed by UV/VIS at 280nm and 410nm absorbance, and size exclusion high performance liquid chromatography (SE-HPLC) to determine antibody concentration and aggregate formation. Upon preliminary testing, acetate was eliminated as a potential storage buffer and an additional 24 full factorial DoE was performed that combined the stabilizing effect of arginine with the buffering capacity of histidine. From this final DoE, a formulation combining 100 mM arginine, 50 mM histidine, and 100 mM NaCl at a pH of 6.5 was identified that substantially improved the stability of the model IgG3 both over time and after multiple freeze/thaw cycles.

Improved stability of a model, difficult to formulate IgG3 by DoE-based evaluation of buffer formulations

249th National Meeting (2015)

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