High binding capacity surface grafted monolithic columns for cation exchange chromatography of proteins and peptides

Item Type Journal Article
Author Jana Krenkova
Author Andrea Gargano
Author Nathan A. Lacher
Author Jeffrey M. Schneiderheinze
Author Frantisek Svec
URL http://www.sciencedirect.com/science/article/B6TG8-4X1J74J-2/2/6920a1a6df442cd39d6fca0a18a87a88
Volume 1216
Issue 40
Pages 6824-6830
Publication Journal of Chromatography A
ISSN 0021-9673
Date Octubre 2, 2009
DOI 10.1016/j.chroma.2009.08.031
Accessed 2009-10-07 11:52:08
Library Catalog ScienceDirect
Abstract Poly(glycidyl methacrylate-co-ethylene methacrylate) monoliths have been prepared in 100 [mu]m i.d. capillaries and their epoxy groups hydrolyzed to obtain poly(2,3-dihydroxypropyl methacrylate-co-ethylene methacrylate) matrix. These polymers were then photografted in a single step with 2-acrylamido-2-methyl-1-propanesulfonic acid and acrylic acid to afford stationary phases for a strong and a weak cation exchange chromatography, respectively. Alternatively, poly(ethylene glycol) methacrylate was used for grafting in the first step in order to enhance hydrophilicity of the support followed by photografting with 2-acrylamido-2-methyl-1-propanesulfonic acid or acrylic acid in the second step. These new columns were used for the separation of proteins and peptides. A mixture of ovalbumin, [alpha]-chymotrypsinogen, cytochrome c, ribonuclease A and lysozyme was used to assess the chromatographic performance for large molecules while a cytochrome c digest served as a model mixture of peptides. All tested columns featured excellent mass transfer as demonstrated with very steep breakthrough curves. The highest binding capacities were found for columns prepared using the two step functionalization. Columns with sulfonic acid functionalities adsorbed up to 21.5 mg/mL lysozyme while the capacity of the weak cation exchange column functionalized with acrylic acid was 29.2 mg/mL.
Title High binding capacity surface grafted monolithic columns for cation exchange chromatography of proteins and peptides
Date Added 2009-10-07 07:52
Date Modified 2009-10-07 07:52

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