High Throughput Formulation Screening for Global Aggregation Behaviors of Three Monoclonal Antibodies

Global aggregation behaviors of three distinct monoclonal antibodies were characterized by high throughput, multiassay analysis. First, extensive screening of formulations was performed using both incubation at elevated temperature and differential thermal scanning. In incubation studies, formulation conditions representing native favored, native favored but with particulate formation, unfolding with slow aggregation, and fast aggregation with or without phase separation were mapped across a wide range of pH and ionic strength. The sample types or aggregation kinetic scenarios were classified based on fluorescence spectroscopy, light scattering, and micron particle count. Furthermore, apparent melting point was determined for each formulation condition by differential thermal scanning. The global aggregation behaviors and their apparent melting points together highlight the common underlying aggregation pathways and kinetics for the three antibodies. Overall, incorporating multistage aggregation mechanisms in multivariate data analysis provides valuable insights to what and how high throughput techniques can be implemented. Understanding global aggregation behaviors is a key element toward development of rational screening approach. © 2011 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 100:2120-2135, 2011