Advances in recombinant DNA biotechnology have stimulated demand for the development of protein formulations within the pharmaceutical industry. A major challenge in preparation of protein formulations is their often poor stability during formulation processing and storage. Combinations of analytical techniques that probe different aspects of protein structure are often required to ensure that higher order (secondary and tertiary) structure and biological activity are maintained. Vibrational spectroscopy has received increasing attention as it offers effective, reliable, fast, simple and non-destructive measurement of protein structure. This review describes recent applications of near-infrared, infrared and Raman spectroscopy for protein analysis. This paper is intended to serve as guidance for researchers beginning in spectral analysis. It is hoped that the information herein will encourage and facilitate research associated with the use of these spectroscopic techniques in protein pharmaceutics.