433864 Rationally Engineering Post-Translational Protein Pegylation for Improved Stability and Function

Wednesday, November 11, 2015: 5:03 PM
151A/B (Salt Palace Convention Center)
Kristen M. Wilding and Bradley C. Bundy, Chemical Engineering, Brigham Young University, Provo, UT

Years of work conjugating the biocompatible polymer polyethylene glycol (PEG) to proteins have already resulted in the development of FDA-approved PEGylated protein therapeutics that are more stable and have longer functional lifetimes than their un-PEGylated counterparts. The post-translational covalent attachment of PEG groups, or PEGylation, increases a drug’s molecular weight, thereby reducing clearance from the body. PEGylation also shields the protein from proteases and can thermodynamically stabilize a protein’s conformation. Polymer-protein conjugation is a challenging task, however; polymer additions may interfere with protein folding and disrupt the protein or enzyme function. Thus, the attachment site of PEG on a protein must be optimized to achieve maximum stability and activity. Using cell-free protein synthesis and the incorporation of non-canonical amino acids, we examine the optimal conditions for conjugating PEG to proteins and compare the effect of PEGylation at different sites. This work will provide a basis for the development of structure-based guidelines to aid in the rational selection of optimal PEGylation sites.

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