Aldehyde PEGylation Kinetics: A Standard Protein versus a Pharmaceutically Relevant Single Chain Variable Fragment

作者：Moosmann Anna^*; Blath Jessica; Lindner Robert; Mueller Egbert; Boettingert Heiner

来源：Bioconjugate Chemistry, 2011, 22(8): 1545-1558.

DOI：10.1021/bc200090x

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.

• 出版日期2011-8