Activation of Sepharose with epichlorohydrin in water-free system for preparation of high capacity immobilized metal ion affinity adsorbent

Song Ping Zhang, Rui Ding, Zhi Guo Su, Ping Wang

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Epoxy activation of agarose gel Sepharose 6 FF by epichlorohydrin (ECH) was carried out in a water-free medium with dimethyl sulfoxide as solvent. The activated agarose gel was subsequently coupled with iminodiacetic acid and Cu2+ ions for preparation of immobilized metal ions affinity chromatographic (IMAC) adsorbent. The results indicated that the epoxy activation efficiency was significantly enhanced in the water-free system, such that a epoxy density of agarose gels as high as 165 μmol/mL was achieved at the optimized conditions of 40%(ψ) ECH, 0.02 g/mL NaOH, 50°C and 4 h, which is about 50% higher than the highest value reported so far. The prepared IMAC adsorbent showed a high chelating capacity for Cu2+ ion up to 128.3 mol/mL, thus it ensured a high equilibrium adsorption capacity of 2.05 mmol/L for bovine serum albumin, and 90% of the adsorbed protein could be efficiently eluted when 0.5 mol/L imidazole was used as elute.

Original languageEnglish (US)
Pages (from-to)971-975
Number of pages5
JournalGuocheng Gongcheng Xuebao/The Chinese Journal of Process Engineering
Volume10
Issue number5
StatePublished - Oct 1 2010

Keywords

  • Activation
  • Agarose gel
  • Epichlorohydrin
  • Immobilized metal affinity chromatography

Fingerprint Dive into the research topics of 'Activation of Sepharose with epichlorohydrin in water-free system for preparation of high capacity immobilized metal ion affinity adsorbent'. Together they form a unique fingerprint.

Cite this