BIOT 10 |
| Purpose The purpose of this research was to determine the aggregation induced by adsorption of a monoclonal antibody to various microparticle surfaces relevant to the final fill, finish and storage of a biopharmaceutical product. We also investigated how formulation excipients can either aggravate or mitigate microparticle induced aggregation. Methods Adsorption was performed by incubation of IgG (antistreptavidin, donated by Amgen Inc.) with microparticles. The suspensions were centrifuged and the protein in the supernatant quantified by UV at 280 nm or SEC and UV at 280 nm. Adsorbed and aggregated protein was then determined by mass balance. Results The aggregation of IgG induced by adsorption was found to be strongly affected by both the surface type and the solution conditions. Tungsten microparticles caused dramatic IgG loss from solution. Addition of 0.01% Tween 20® to the buffer stabilized IgG from silica induced aggregation, but the addition of sodium chloride increased the observed aggregation. Conclusions Protein aggregation is increased in the presence of microparticles; however, solution conditions can dramatically alter the rate and extent of induced aggregation. Incubation studies with microparticles could be used for accelerated screening of formulation stability with respect to adsorption induced aggregation. |
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Biophysical and Biomolecular Symposium: Protein Stability
8:00 AM-11:10 AM, Sunday, August 19, 2007 BCEC -- 107 A/B, Oral
Division of Biochemical Technology |