Purification of PEGylated Protein Using Membrane Chromatography

N-terminus-specific PEGylation was used to produce mono-PEGylated lysozyme However, some di- and tri-PEGylated proteins were also produced due to side chain reaction The reaction products were characterized by chromatographic and electrophoretic methods. Commercial cation exchange membrane Sartobind S was used for chromatographic purification of PEGylated lysozyme, the basis of separation being the shielding of protein charge by PEG Using the membrane chromatographic method, lysozyme and mono-, di-, and tri-PEGylated lysozyme could be resolved into separate peaks. Increasing the superficial velocity during chromatographic separation from 24 cm/h to 240 cm/h increased both protein binding capacity and resolution due to enhancement of protein mass transfer coefficient (C) 2010 Wiley-Liss, Inc and the American Pharmacists Association J Pharm Sci 99 3326-3333 2010