Related Experiment Video
Updated: Jun 18, 2026

Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography
Published on: January 15, 2014
Analytical and preparative separation of PEGylated lysozyme for the characterization of chromatography media
Anna Moosmann1, Jessica Christel, Heiner Boettinger
1Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. anna.moosmann@izi.uni-stuttgart.de
Abstract:
The effect of PEGylation on cation exchange chromatography was studied with poly(ethylene glycol) of different chain lengths (5kDa, 10kDa and 30kDa) using lysozyme as a model system. A stable binding via reduction of a Schiff base was formed during random PEGylation on lysine residues with methoxy-PEG-aldehyde. A purification method for PEGylated proteins using cation exchange chromatography was developed, and different isoforms of mono-PEGylated lysozyme were isolated. TSKgel SP-5PW and Toyopearl GigaCap S-650M showed the best performance of all tested cation exchange resins, and the separation of PEGylated lysozyme could be also scaled up to semi-preparative level. Size-exclusion chromatography, SDS-PAGE and MALDI-TOF mass spectrometry were used for analysis. Separated mono-PEGylated lysozyme of different sizes was used to determine dynamic binding capacities (DBC) and selectivity of cation exchange chromatography resins. An optimization of binding conditions resulted in a more than 20-fold increase of DBC for Toyopearl GigaCap S-650M with 30kDa mono-PEGylated lysozyme.
Related Concept Videos
Types Of Column Chromatography
Gel Filtration Chromatography
When the...
Affinity Chromatography
Capillary Electrophoresis: Applications
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...

