%0 Journal Article %1 Moosmann2010 %A Moosmann, Anna %A Christel, Jessica %A Boettinger, Heiner %A Mueller, Egbert %D 2010 %J Journal of Chromatography A %K 2010 boettinger izi %N 2 %P 209--215 %R 10.1016/j.chroma.2009.11.031 %T Analytical and preparative separation of PEGylated lysozyme for the characterization of chromatography media %U https://www.ncbi.nlm.nih.gov/pubmed/19963219 %V 1217 %X The effect of PEGylation on cation exchange chromatography was studied with poly(ethylene glycol) of different chain lengths (5 kDa, 10 kDa and 30 kDa) 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 30 kDa mono-PEGylated lysozyme. © 2009 Elsevier B.V. All rights reserved. %Z Moosmann, AnnaChristel, JessicaBoettinger, HeinerMueller, EgbertengResearch Support, Non-U.S. Gov'tNetherlandsJ Chromatogr A. 2010 Jan 8;1217(2):209-15. doi: 10.1016/j.chroma.2009.11.031. Epub 2009 Nov 17. %7 2009/12/08 %@ 0021-9673

@article{Moosmann2010, abstract = {The effect of PEGylation on cation exchange chromatography was studied with poly(ethylene glycol) of different chain lengths (5 kDa, 10 kDa and 30 kDa) 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 30 kDa mono-PEGylated lysozyme. {\textcopyright} 2009 Elsevier B.V. All rights reserved.}, added-at = {2018-02-01T15:54:10.000+0100}, annote = {Moosmann, AnnaChristel, JessicaBoettinger, HeinerMueller, EgbertengResearch Support, Non-U.S. Gov'tNetherlandsJ Chromatogr A. 2010 Jan 8;1217(2):209-15. doi: 10.1016/j.chroma.2009.11.031. Epub 2009 Nov 17.}, author = {Moosmann, Anna and Christel, Jessica and Boettinger, Heiner and Mueller, Egbert}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2bf9d65b063ea4529a01d908875c5065f/cristiano}, doi = {10.1016/j.chroma.2009.11.031}, edition = {2009/12/08}, interhash = {a775567e743188c0dbc1821f8878e1b6}, intrahash = {bf9d65b063ea4529a01d908875c5065f}, isbn = {0021-9673}, issn = {00219673}, journal = {Journal of Chromatography A}, keywords = {2010 boettinger izi}, number = 2, pages = {209--215}, pmid = {19963219}, timestamp = {2018-07-25T12:32:54.000+0200}, title = {{Analytical and preparative separation of PEGylated lysozyme for the characterization of chromatography media}}, url = {https://www.ncbi.nlm.nih.gov/pubmed/19963219}, volume = 1217, year = 2010 }