Strong cation-exchange chromatography of proteins on a sulfoalkylated monolithic cryogel

Percin I., KHALAF R., BRAND B., MORBIDELLI M., GEZICI O.

JOURNAL OF CHROMATOGRAPHY A, cilt.1386, ss.13-21, 2015 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 1386
  • Basım Tarihi: 2015
  • Doi Numarası: 10.1016/j.chroma.2015.01.075
  • Dergi Adı: JOURNAL OF CHROMATOGRAPHY A
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.13-21
  • Anahtar Kelimeler: Gradient elution, HPLC, Ion-exchange, Protein, Separation, Yamamoto Model, GRAFT-POLYMERIZATION, OPERATING-CONDITIONS, ELUTION CURVES, DIRECT CAPTURE, COLUMN, PERFORMANCE, PREDICTION, RETENTION, MATRICES
  • Hacettepe Üniversitesi Adresli: Evet

Özet

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 glycidyl-methacrylate (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) N-2 adsorption, and used as a stationary phase for strong-cation exchange chromatography of some proteins, such as a-chymotrypsinogen, cytochrome c and lysozyme. Water permeability of the column was calculated according to Darcy's law (2.66 x 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. (c) 2015 Elsevier B.V. All rights reserved.