Estimating BSA Aggregation Kinetics with Dynamic Light Scattering and Raman Spectroscopy
White Paper Apr 13, 2016
Malvern Instruments Limited
Dynamic light scattering (DLS) is a widely accepted technique for characterizing the hydrodynamic radius of nano-scale materials in suspension. The application of DLS measurements to the analysis of protein therapeutics has increased dramatically over the last ten years and the technique is now considered a staple of analytical technology in the biopharmaceutical sector. The ability of DLS to measure formulationlevel concentrations, coupled with its inherent sensitivity to large particles, make it the perfect tool for the study of protein aggregation in solution.
Raman spectroscopy is capable of delivering detailed structural information about protein formulations, through access to secondary and tertiary structural markers. Band intensity changes, broadening, and shifts can all be directly assigned to protein structural changes as a function of stress and environment. Such structural changes are often precursors to aggregation, and can be probed directly with Raman spectroscopy.
The combination of DLS and Raman spectroscopy offers the ability to measure changes in size concurrent with changes in molecular structure. This allows the balance between protein aggregation and protein structural changes to be investigated as a function of changing formulation conditions. Rate constants for the underlying aggregation and structural changes can be derived directly from the resulting data.
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