title: Aldehyde PEGylation kinetics: a standard protein versus a pharmaceutically relevant single chain variable fragment.
abstract: The mPEG-aldehyde PEGylation with two different PEG sizes and two proteins was experimentally determined with respect to yield, conversion, and selectivity. The kinetic behavior of these PEGylation reactions was simulated using a numerically solved set of differential equations. We show that the assumption of an inactivation of mPEG-aldehyde is crucial for the simulation of the overall PEGylation and that the inactivation is pH-dependent. We further demonstrate that ideal PEGylation parameters such as pH, temperature, reaction time, and protein concentration need to be chosen carefully depending on the protein and PEG size. In terms of selectivity and yield, we show that the reaction should be stopped before the highest mono-PEG concentration is reached. Moreover, room temperature and a slightly acidic pH of approximately 6 are good starting points. In conclusion, selectivity can be optimized choosing a shorter reaction time and a reduced reaction temperature.
authors.affiliation: Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. anna.moosmann@izi.uni-stuttgart.de
authors.forename: Anna
authors.initials: A
authors.lastname: Moosmann
authors.forename: Jessica
authors.initials: J
authors.lastname: Blath
authors.forename: Robert
authors.initials: R
authors.lastname: Lindner
authors.forename: Egbert
authors.initials: E
authors.lastname: Müller
authors.forename: Heiner
authors.initials: H
authors.lastname: Böttinger
mesh_terms: D000447:Aldehydes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008962:Models, Theoretical; D011092:Polyethylene Glycols / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D012015:Reference Standards; D057127:Single-Chain Antibodies / Q000737:chemistry; D013696:Temperature
chemical_list: D000447:Aldehydes; D011506:Proteins; D057127:Single-Chain Antibodies; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol