Model-based development of an on-column PEGylation process. Issue 2 (4th November 2015)

• Record Type: Journal Article
• Authors: Pfister, David; Ingold, Oliver; Morbidelli, Massimo
• Is Part Of: Reaction chemistry & engineering. Volume 1:Issue 2(2016)
• Page Start: 204
• Page End: 217
• Publication Date: 2015-11-04
• Subjects: Reaction mechanisms (Chemistry) -- Periodicals; Chemical engineering -- Periodicals; Chemical engineering; Reaction mechanisms (Chemistry); Periodicals; 547.705
• DOI: 10.1039/c5re00019j
• Languages: English
• ISSNs: 2058-9883

Abstract

Abstract : On-column PEGylation appears as an interesting alternative to classical solution reaction for more selective synthesis of the targeted mono-PEGylated protein. Indeed, it has the potential to inhibit the formation of the multi-PEGylated species and provide site selectivity by restricting the coupling reaction to fewer reaction sites. Abstract : Considering the high production cost of therapeutic proteins, post-translational modifications such as PEGylation have to be highly efficient and optimized so as not to waste the valuable protein to be modified. Among the different processes available, on-column PEGylation appears as an interesting alternative to the classical solution reaction for more selective synthesis of the targeted mono-PEGylated protein since it has the potential to restrict the coupling reaction to fewer reaction sites. In this work, the on-column PEGylation of α-lactalbumin using a strong anion-exchange resin is investigated. The influence of pH, reaction time, PEG to protein ratio and protein load is investigated. The on-column PEGylation process is shown to be more selective for lower degrees of PEGylation than the classical solution PEGylation reaction. Finally, the experimental results are described by a mathematical model in order to gain a more complete understanding of the mechanisms involved and develop a tool for process design and development.