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

作者：Lihme Allan^*; Hansen Marie Bendix; Andersen Inga Vaarst; Burnouf Thierry

来源：Analytical Biochemistry, 2010, 399(1): 102-109.

DOI：10.1016/j.ab.2009.12.002

• Plasma
• Fractionation
• Expanded bed chromatography
• Prothrombin complex
• Factor VIII
• von Willebrand factor
• Immunoglobulins
• alpha 1-Antitrypsin
• Albumin

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 mu m) 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) (similar to 1 IU/ml and 0.6 IU/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 alpha 1-anti-trypsin 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.

• 出版日期2010-4-1