| The ability to successfully formulate and manufacture therapeutic protein dosage forms requires a thorough understanding of their physico-chemical properties. Proteins are inherently dynamic molecules of marginal stability. These properties present unique challenges to the pharmaceutical scientist attempting to develop protein based therapeutics. The physicochemical stability and biological functions of proteins are thought to be intimately related to their global flexibility, intramolecular fluctuations and various other dynamic processes. Our understanding of these relationships, however, is incomplete but undeniably necessary for the development of efficacious therapies. Therefore, a better understanding of the complex inter-relationships between protein flexibility and stability should enable the rational design and optimization of protein formulation conditions based on protein dynamics.; Multiple experimental methods were employed that are sensitive to a wide range of protein dynamic processes to help elucidate the molecular determinants of protein stability. To this end, various external perturbations were used to modulate both protein stability and flexibility. This work provides evidence that previously hypothesized relationships between protein dynamic behavior and stability are inadequate. Furthermore, evidence supporting a much more complex inter-relationship between the coupling of both flexible and rigid regions to stability is established. Although a single dynamic processes controlling protein stability was not clearly determined, multiple factors that contribute to stability are discussed. |
