Add time:08/29/2019         Source:sciencedirect.com

A critical challenge of PEGylation is the production of the desired PEGylated protein form at a high yield. In this study, a kinetic model was constructed successfully to describe the PEGylation reaction of recombinant hirudin variant-2 (HV2) with monomethoxy-PEG-succinimidyl carbonate (mPEG-SC) by fitting the experimental data. Moreover, PEGylation reaction conditions were investigated using the established model and the corresponding experiments to determine the optimal condition to achieve the mono-PEG-HV2 at the desired yield. The model predictions agreed well with the experimental data. Several important process parameters (maximum theoretical yield of mono-PEG-HV2 (ymax), critical PEG/HV2 molar ratio (mcrit) and reaction time to achieve ymax (tmax)) and their mathematical equations were obtained to determine the optimum reaction conditions. Among reaction conditions affecting the PEGylation rates, pH and temperature displayed little effect on ymax, but ymax increased as PEG size increased. Optimal reaction condition to produce mono-PEG-HV2 was as follows: pH and temperature could vary in a certain range; whereas PEG/HV2 molar ratio should be slightly greater than mcrit and the reaction should be stopped at tmax. The results of this study indicate that the proposed reaction kinetic model can provide a possible mechanism interpretation for real PEGylation reactions and optimize efficiently the PEGylation step.

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