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

Lihme A., Hansen M., Andersen IV., Burnouf T. Anal. Biochem. 2010; 399(1):102-9.

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 d(v)(0.5)=190 and 37 microm) to isolate at 50 to 80% recovery a fraction containing 4 to 7 IU/ml factor II (FII), factor IX (FIX), and factor X (FX) (specific activity >1 IU/mg) and another enriched in FVIII and von Willebrand factor (vWF) (approximately 1 IU/ml and 0.6 IU/mg, respectively).  The flow-through was assorbed 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 alpha1-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.

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