Abstract
This chapter describes applications of attenuated total reflection (ATR)–Fourier transform infrared (FTIR) spectroscopy and ATR-FTIR spectroscopic imaging to studies of proteins. The latter method combines ATR-FTIR spectroscopy with an infrared array detector for obtaining both spatial and chemical information from protein samples. Two imaging modes, micro- and macro-ATR, provide a range of imaging fields of view and spatial resolutions. Micro-ATR-FTIR imaging has been successfully used to study hanging drop protein crystallization with high spatial resolution imaging, while macro-ATR-FTIR imaging provided new opportunities for in situ studies of protein crystallization and aggregation, as well as the effect of different wettability surface properties on protein adsorption and crystallization. Recent pioneering applications of ATR-FTIR spectroscopy involve analysis of protein ligand denaturation on chromatography columns, critical for improved industrial purification of biotherapeutics. These and other new high-throughput chemical imaging approaches are discussed in this chapter.
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