article accepted in Methods Mol Biol
Purification of PEGylated Proteins, with the Example of PEGylated Lysozyme and PEGylated scFv
Anna Moosmann 1, Egbert Müller, and Heiner Böttinger
1Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany, an-na-mo@gmx.de.
Abstract :
PEGylation is a common and highly accepted possibility for half-life prolongation of proteins by increasing the hydrodynamic size. The chromatographic purification of PEGylated protein, using PEG (poly-ethylene glycol) of different PEG chain lengths, with the example of lysozyme and a scFv, is described in detail here, and helpful suggestions for the purification of other PEGylated proteins are listed. The relevant characterization methods for PEGylated proteins, important for the successful purification, are also described. The purification starts with a CEX (cation exchange) chromatography leading to about 95 % purity for polishing HIC (hydrophobic interaction chromatography) is described.