Protein resolution in elution chromatography using novel cation-exchange polymer-brush-immobilized particles

Takato Harayama, Yusuke Okamura, Yuichi Shimoda, Daisuke Umeno, Kyoichi Saito, Naoyuki Shinohara, Noboru Kubota

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

This study investigates improvements in high-resolution elution chromatography of proteins loaded to a bed charged by way of cation-exchange polymer-brush-immobilized particles. A sulfonic acid group as a cation-exchange group was introduced into a polymer brush grafted onto a polyethylene particle with an average diameter of 35 μm. A bed charged with the resulting cation-exchange polymer-brush-immobilized particles, SS bed, has a sulfonic acid group density of 0.36 mmol/mL-bed. A mixture of α-chymotripsinogen A (chy), cytochrome c (cyt), and lysozyme (lys) was loaded onto the SS bed. The subsequent linear gradient elution performance of the proteins adsorbed by the SS bed with 20 mM phosphate buffer (pH 6.0) and 1 M NaCl is compared with those by commercially available cation-exchange-bead-packed beds. The resolutions of the SS bed of both the chy/cyt and cyt/lys pairs at a linear velocity of 600 cm/h exceeded 1.9 at a loading amount of 0.8 mg/mL-bed, which was not achieved using the beds charged with SOURCE® 30S, POROS® 50HS, or Fractogel® EMD SO3- (s). Even at a loading amount of 12 mg/mL-bed, the resolutions of the SS bed for the chy/cyt and cyt/lys pairs at a linear velocity of 300 cm/h were 1.4 and 2.7, respectively.

Original languageEnglish
Pages (from-to)896-902
Number of pages7
JournalJOURNAL OF CHEMICAL ENGINEERING OF JAPAN
Volume45
Issue number11
DOIs
Publication statusPublished - 2012
Externally publishedYes

Keywords

  • Cation Exchange
  • Elution Chromatography
  • Polymer Brush
  • Protein Purification
  • Radiation-Induced Graft Polymerization

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

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