SHANNON SOUTHALL, CHARLENE E. BRISBANE, AMOL KETKAR
Biopharmaceutical Technologies, Biopharmaceutical Development, GlaxoSmithKline Pharmaceutical R&D, King of Prussia PA, USA
Posted with permission from Shannon Southall
Purpose:
The objectives of this study are: a) to highlight the importance of sub-visible particle analysis during formulation development, and b) to evaluate the ability of common formulation excipients to protect against particle formation.
Methods:
Different mAb formulations were subjected to freeze/thaw and shear, stresses that are known to cause sub-visible particle formation in protein solutions. The effects of buffer, surfactant, amino acid, chelating agent, and sugar excipients were evaluated in terms of ability to stabilize the protein against these stresses and protect against the formation of particles. Micro-flow imaging (MFI) and light obscuration were used to assess particle counts, size range, and morphology. Additionally, the impact of these stresses on other critical quality attributes was evaluated by standard biochemical analyses.
Results:
Both shear and freeze-thaw stress resulted in sub-visible particle formation, particularly in the <10 micron range where an increase of more than 500 fold was observed. Despite this significant rise in sub-visible particles, the level of soluble aggregates (as measured by SEC) was not affected, and all other product quality attributes were retained.
The use of surfactant was most effective in preventing particle formation. This is likely due to the type of stresses chosen in this study which cause particle formation via interfacial destabilization.
In general, MFI was found to be a more sensitive method to assess particle formation as compared to light obscuration. Particle counts observed by MFI were at least three times higher than those detected by light obscuration, although the trends observed by the two techniques were similar
Conclusion:
Particle analysis is an important component of formulation development and should be used in conjunction with other biochemical assays to identify the optimal formulation.
PRESENTED AT:
2010 AAPS National Biotechnology Conference, San Francisco CA, USA
2010 AAPS NBC_GSK_Particle Analysis as a Formulation Development Tool.pdf
2010 AAPS NBC GSK Particle Analysis