In a recent paper we have reported the use of hydrophilized polyvinylidene fluoride (PVDF) membrane as environment-responsive stationary phase for the fractionation of unmodified lysozyme and its different PEGylated forms (i.e. mono-, di- and tri-PEGylated lysozyme) [1]. In the current study, a similar hydrophobic interaction based technique is applied to purify mono-PEGylated human serum albumin (HSA) prepared by N-terminus PEGylation reaction of the protein with m-PEG propionaldehyde. This reaction results in the formation of mainly mono-, and di-PEGylated and smaller amounts of tri-PEGylated HSA. The reaction mixture is separated at both analytical and preparative scales. The fractionated samples are analyzed and characterized using techniques such as SDS-PAGE and 2-D electrophoresis and multiple angle light scattering (or MALS) technique. References1. Deqiang Yu, Xiaojiao Shang, Raja Ghosh, Fractionation of different PEGylated forms of a protein by chromatography using environment-responsive membranes. Journal of Chromatography A, 1217 (2010) 5595-5601.

Purification of PEGylated protein by hydrophobic interaction membrane chromatography

241st National Meeting (2011)

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