The Utility Of Mass Spectrometry Based Structural Proteomics In Biopharmaceutical Biologics Development

Proteins are essential components of living organisms and participate in nearly every biochemical process within cells. Examples of the processes include enzyme catalysis, cell signalling, host defense, metabolism, etc. These large, complex bio-molecules are connected by long chains of amino acids, that fold in very intricate patterns giving rise to a unique three-dimensional conformation. The biological function and physicochemical properties of a protein are determined by this higher order structure Ecroyd & Carver (2008); Hegyi & Gerstein (1999); Sadowski & Jones (2009). Most proteins tend to achieve the lowest possible free energy of the polypeptide chain and the surrounding solvent forming a native structure under physiological conditions Anfinsen (1973). This tightly folded conformation typically represents the biologically active state necessary for performing the required biochemical task. However, these macromolecules also have a temporal behavior leading to significant flexibility and dynamic motion because of the fluctuations in the surrounding electrostatic forces and hydrogen bonds that are important for maintaining conformations Henzler-Wildman K Teilum et al. (2009). These temporal variations are important for certain functions such as protein-protein interactions and protein stability Kamerzell & Middaugh (2008); Travaglini-Allocatelli et al. (2009); van der Kamp et al. (2010). Thus, it is the presence of both spatial and temporal characteristics that allows for an ensemble of various molecular conformations to exist in solution. Change in environment of proteins, such as solvent acidity, urea concentration, temperature fluctuations, can change the folding pattern of the protein. Studying these partially or fully denatured states provides insights for understanding a variety of in vivo processes such as structural changes associated with aggregation, signal transduction, and transportation across membranes. Certain biological conditions can cause misfolding and aggregation of proteins, often causing severe disorders such as Alzheimer’s disease, spongiform encephalopathies, and certain forms of diabetes Dobson (2003). Many genetic diseases are caused by protein-folding disorders, because an altered gene results in amodified protein sequence which is not able to undergo native folding and results in the disease phenotype Dobson (2001). Proteins have the ability to interact with one another, and can also bind to smaller ligands, which forms the basis of signaling and regulatory processes, playing a critical role in the mechanisms of drug activity. Owing to the The Utility of Mass Spectrometry Based Structural Proteomics in Biopharmaceutical Biologics Development