title: Strong cation-exchange chromatography of proteins on a sulfoalkylated monolithic cryogel.
abstract: A new strong cation exchanger (SCX) monolithic column was synthesized by at-line surface modification of a cryogel prepared by copolymerization of 2-hydroxyethylmethacrylate (HEMA) and glycidylmethacrylate (GMA). Sodium salt of 3-Mercaptopropane sulfonic acid (3-MPS) was used as the ligand to transform the surface of the monolith into a strong cation exchanger. The obtained material was characterized in terms of elemental analysis, infrared spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Brunauer-Emmett-Teller (BET) N2 adsorption, and used as a stationary phase for strong-cation exchange chromatography of some proteins, such as α-chymotrypsinogen, cytochrome c and lysozyme. Water permeability of the column was calculated according to Darcy's law (2.66×10(-13)m(2)). The performance of the monolithic cryogel column was evaluated on the basis of Height Equivalent to a Theoretical Plate (HETP). Retention behavior of the studied proteins was modeled on the basis of Yamamoto model to understand the role of the ion-exchange mechanism in retention behaviors. The considered proteins were successfully separated, and the obtained chromatogram was compared with that obtained with a non-functionalized cryogel column.
authors: ETH Zurich, Institute for Chemical and Bioengineering, Vladimir-Prelog-Weg 1, CH-8093 Zurich, Switzerland; Hacettepe University, Department of Biology, Molecular Biology Division, Beytepe, 06800 Ankara, Turkey.
authors: Işık
authors: I
authors: Perçin
authors: ETH Zurich, Institute for Chemical and Bioengineering, Vladimir-Prelog-Weg 1, CH-8093 Zurich, Switzerland.
authors: Rushd
authors: R
authors: Khalaf
authors: ETH Zurich, Institute for Chemical and Bioengineering, Vladimir-Prelog-Weg 1, CH-8093 Zurich, Switzerland.
authors: Bastian
authors: B
authors: Brand
authors: ETH Zurich, Institute for Chemical and Bioengineering, Vladimir-Prelog-Weg 1, CH-8093 Zurich, Switzerland.
authors: Massimo
authors: M
authors: Morbidelli
authors: ETH Zurich, Institute for Chemical and Bioengineering, Vladimir-Prelog-Weg 1, CH-8093 Zurich, Switzerland; Nigde University, Faculty of Science and Arts, Chemistry Department, 51200, Nigde, Turkey. Electronic address: ogezici@gmail.com.
authors: Orhan
authors: O
authors: Gezici
mesh_terms: D000327:Adsorption; D002411:Cation Exchange Resins / Q000032:analysis; D002412:Cations / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D002919:Chymotrypsinogen / Q000737:chemistry / Q000302:isolation & purification; D059985:Cryogels / Q000737:chemistry*; D045304:Cytochromes c / Q000737:chemistry / Q000302:isolation & purification; D008689:Methacrylates / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D017550:Spectroscopy, Fourier Transform Infrared; D013451:Sulfonic Acids / Q000737:chemistry*
chemical_list: D002411:Cation Exchange Resins; D002412:Cations; D059985:Cryogels; D008689:Methacrylates; D011506:Proteins; D013451:Sulfonic Acids; C005044:hydroxyethyl methacrylate; D045304:Cytochromes c; D002919:Chymotrypsinogen; D009113:Muramidase
keywords: Gradient elution; HPLC; Ion-exchange; Protein; Separation; Yamamoto Model