Do you currently experience a lack of reliable methods for the quantification of submicron (0.1 to 1 mm) protein aggregates? Would you like to learn more about how field-flow fractionation and light scattering are suitable techniques for studying aggregates and supramolecular assemblies? If so, this presentation from Separation Science, in collaboration with Wyatt Technology, can help you achieve your goal.
Protein aggregation in biotherapeutic drugs is a major concern as they affect the effective dosage and may cause immunogenic responses in patients. However, a lack of reliable analytical methods has hindered the quantification of submicron (0.1 to 1 mm) protein aggregates and a detailed understanding of their formation kinetics. In this study, a simple asymmetrical flow field-flow fractionation (AF4) method is developed and used to investigate nanometer (<0.1 mm) and submicron (0.1-1 mm) aggregates of heat-stressed anti-streptavidin (anti-SA) IgG1. The Lumry-Eyring nucleated polymerization model for non-native protein aggregation is fit to AF4 data and kinetic analysis is performed to identify the mechanism of aggregate formation. This work show cases the importance of a separation technique when studying complex and dynamic systems. Questions about the impact of a separation method on potentially ‘labile’ analytes are also be addressed.
By viewing this presentation you will learn about...
- The field-flow fractionation separation mechanism
- The combination of FFF with light scattering, which provides an enhanced understanding of (complex) analyte systems
- Thinking about dilution effects – it may not be as simple or as bad as you think.
Kim R. Williams is a Professor of Chemistry at the Colorado School of Mines. Before joining the faculty at CSM in 1997, she worked with the late Professor J. Calvin Giddings as a postdoctoral fellow and then in the dual role of Assistant Director of the Field-Flow Fractionation Research Center and Adjunct Professor of Chemistry at the University of Utah. She holds a B.Sc. in Chemistry from McGill University and a Ph.D. in Chemistry from Michigan State University. (Williams aka Ratanathanawongs was born and raised in Bangkok, Thailand.)