Circular Dichroism Spectroscopy for Structural Characterization of Proteins
The large size, physicochemical complexity and rather labile nature of proteins present significant challenges in delivery of pharmaceutical proteins. First, the native structure of a given protein drug molecule must be characterized in order to define the active compound. Next, for the successful development of a formulation or drug delivery system for said molecule, compatibility of the system with the protein should be demonstrated. This is a challenging task as formulations and delivery systems often contain chromophores or particulates that absorb or scatter light, preventing the direct use of several of the most commonly used techniques for structural characterization.
The single most important spectroscopic method for the rapid elucidation of protein structure in solution is circular dichroism (CD) spectroscopy. In this chapter, an overview of the method is given, with a discussion of the instrumentation, several specific examples of applications and general experimental recommendations for CD experiments, including a special perspective on challenging samples as may be encountered in pharmaceutical sciences.
KeywordsCircular dichroism (CD) Synchrotron radiation CD Secondary, tertiary and quaternary structure Protein folding
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