Ubiquitous ultraviolet absorption spectroscopy, despite the availability of more sophisticated techniques, remains an indispensable tool that can give an initial insight into the concentration and aggregation state of protein samples. The high degree of reproducibility afforded by diode-array spectrophotometers, combined with their powerful vendor-supplied algorithms, presents an opportunity for improving the accuracy and throughput for their use in pharmaceutical development. In this review, factors important to optimal utilization of the technique, as applied to the development of monoclonal antibodies, are discussed, and specific methodologies are described. In particular, techniques to probe the intrinsic structural properties of proteins, and their behavior under a wide variety of conditions, through the application of second-derivative spectroscopy combined with advanced computational treatments are presented. The information contained in this review is specifically directed to practitioners of the technique in contemporary research and development settings.
Keywords: UV-spectroscopy; physicochemical properties; protein formulation; protein structure; solubility; stability.
Copyright © 2010 Wiley-Liss, Inc.