Analytical Ultracentrifugation (AUC) is one of the most powerful biophysical tools used today for the characterization of biological samples ranging from small drug molecules to intact viruses, vesicles and microparticles. AUC works with biological samples in the native state and does not depend on a reporter species or custom-coated substrates which are required by techniques such as NMR, EPR, SPR and fluorescence spectroscopy. AUC separates biomolecules based upon both molecular mass and anisotropy, unlike Size Exclusion Chromatography (SEC), which separates molecules based only on their hydrodynamic volume. Furthermore, AUC is not just useful for characterizing the oligomerization/aggregation state of biological samples, but also to quantify interactions between different species.
In this talk, we will start with the principles of analytical ultracentrifugation and take a tour through the technology behind modern AUC, including UV/Vis absorbance mode and interference mode signal detection, and look at experiment design and setup. Following, we will address the different types of AUC experiments (equilibrium and velocity), compare and contrast their merits with sample data, and touch upon the principles of data processing (to be covered in-depth in a subsequent webinar). Finally, we will explore a variety of applications with a focus on the unique advantages that AUC brings to the study of various biotherapeutics, polymers, nanoparticles, and others – and how AUC compares to and complements other analytical techniques.
Learning Objectives:
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