%0 Journal Article %1 Moosmann2012 %A Moosmann, Anna %A Gerlach, Elke %A Lindner, Robert %A Böttinger, Heiner %D 2012 %J Journal of Chromatography A %K 2012 boettinger izi %P 90--96 %R 10.1016/j.chroma.2012.03.004 %T Purification of a PEGylated single chain Fv %U http://www.ncbi.nlm.nih.gov/pubmed/22444429 %V 1236 %X In this manuscript we describe the two-step purification of a mono-PEGylated anti-epidermal growth factor receptor (EGFR) single-chain Fv. A weak cation exchanger was used for capture. Elution using arginine suppressed protein aggregation and allowed a very good resolution with purity and product-recovery was above 90\%. Free PEG was removed completely. The use of hydrophobic interaction chromatography (HIC) increased purity to 98\%. Increasing the size of PEG from 5 to 30kDa increased retention on HIC and reduced it on cation exchangers. Bioactivity of PEGylated scFv was confirmed by125I based cell tests. Proteins modified with 5kDa PEG showed higher bioactivity than proteins modified with larger PEGs. The combination of cation exchange and HIC provides a rational and effective basis for PEGylated scFv purification. © 2012 Elsevier B.V.

@article{Moosmann2012, abstract = {In this manuscript we describe the two-step purification of a mono-PEGylated anti-epidermal growth factor receptor (EGFR) single-chain Fv. A weak cation exchanger was used for capture. Elution using arginine suppressed protein aggregation and allowed a very good resolution with purity and product-recovery was above 90{\%}. Free PEG was removed completely. The use of hydrophobic interaction chromatography (HIC) increased purity to 98{\%}. Increasing the size of PEG from 5 to 30kDa increased retention on HIC and reduced it on cation exchangers. Bioactivity of PEGylated scFv was confirmed by125I based cell tests. Proteins modified with 5kDa PEG showed higher bioactivity than proteins modified with larger PEGs. The combination of cation exchange and HIC provides a rational and effective basis for PEGylated scFv purification. {\textcopyright} 2012 Elsevier B.V.}, added-at = {2023-06-29T13:07:55.000+0200}, author = {Moosmann, Anna and Gerlach, Elke and Lindner, Robert and B{\"{o}}ttinger, Heiner}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2988ac5648cbba5edd5e58df1081855ed/fabian}, doi = {10.1016/j.chroma.2012.03.004}, interhash = {bcfa17481bff9a2ba392451c3c428007}, intrahash = {988ac5648cbba5edd5e58df1081855ed}, issn = {00219673}, journal = {Journal of Chromatography A}, keywords = {2012 boettinger izi}, month = may, pages = {90--96}, pmid = {22444429}, timestamp = {2023-06-29T13:07:55.000+0200}, title = {{Purification of a PEGylated single chain Fv}}, url = {http://www.ncbi.nlm.nih.gov/pubmed/22444429}, volume = 1236, year = 2012 }