A novel core fractionation process of human plasma by expanded bed adsorption chromatography

Allan Lihme, Marie Bendix Hansen, Inga Vaarst Andersen, Thierry Burnouf

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Current plasma fractionation technology combines ethanol precipitation with packed bed chromatography. We have developed a novel core fractionation process comprising five expanded bed adsorption (EBA) chromatographic steps on high-density modified agarose/tungsten carbide beads. Plasma was first chromatographed on two diethyl amino-ethyl (DEAE)-tungsten carbide agarose adsorbents (respective mean particle diameters of dv(0.5)=190 and 37μm) to isolate at 50 to 80% recovery a fraction containing 4 to 7IU/ml factor II (FII), factor IX (FIX), and factor X (FX) (specific activity >1IU/mg) and another enriched in FVIII and von Willebrand factor (vWF) (∼1IU/ml and 0.6IU/mg, respectively). The flow-through was adsorbed on 4% agarose-10% tungsten carbide beads coupled with an acidic mixed-mode ligand to isolate an 80% pure immunoglobulin G (IgG) at a 93% step recovery. A highly purified α1-antitrypsin was isolated at 95% step recovery by adsorbing the flow-through on 4% epoxy-crosslinked agarose-10% tungsten carbide adsorbent material coupled with a cationic ligand. Isolation of 98% pure albumin was achieved at a 99% step recovery by pH 4.5 adsorption of the flow-through on 6% agarose-10% tungsten carbide beads coupled with an acidic mixed-mode ligand. EBA may represent a feasible alternative core plasma fractionation tool.

Original languageEnglish
Pages (from-to)102-109
Number of pages8
JournalAnalytical Biochemistry
Volume399
Issue number1
DOIs
Publication statusPublished - Apr 2010
Externally publishedYes

Keywords

  • α1-Antitrypsin
  • Albumin
  • Expanded bed chromatography
  • Factor VIII
  • Fractionation
  • Immunoglobulins
  • Plasma
  • Prothrombin complex
  • Von Willebrand factor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

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