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.
Yu
Deqiang
D
Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.
Ghosh
Raja
R
D002138:Calibration; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D008567:Membranes, Artificial; D008970:Molecular Weight; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D011092:Polyethylene Glycols / Q000737:chemistry*; D011312:Pressure; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*
D008567:Membranes, Artificial; D011506:Proteins; D011092:Polyethylene Glycols; D009113:Muramidase