Purification of PEGylated protein using membrane chromatography

✍ Scribed by Deqiang Yu; Raja Ghosh

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 170 KB
• Volume: 99
• Category: Article
• ISSN: 0022-3549
• DOI: 10.1002/jps.22103

✦ Synopsis

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.