A novel matrix for high performance affinity chromatography and its application in the purification of antithrombin III.
Viscose fiber, a regenerated cellulose, was evaluated for using as a novel matrix for high performance affinity chromatography. With a one-step activation with epichlorohydrin, heparin can be readily covalently attached to the matrix. This heparin-viscose fiber material was used for purifying antithrombin III (AT III) from human plasma. The purity of the AT III from this one-step purification is 93% as measured by SDS-PAGE and the protein recovery yield is about 90%. This column is highly specific as described by the dissociation constant of the complex of immobilized heparin and AT III, which was 2.83 x 10(-5)mol/L. And more important, this viscose fiber material demonstrated its excellent mechanical property that allows the flow rate to reach up to 900 cm/h or more.
Zhao
Rui
R
Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Zhongguancun, 100080 Beijing, P.R China. zhaorui@iccas.ac.cn
Luo
Jia
J
Shangguan
Dihua
D
Liu
Guoquan
G
D000990:Antithrombin III / Q000302:isolation & purification*; D002482:Cellulose / Q000648:ultrastructure; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D006493:Heparin / Q000302:isolation & purification; D006801:Humans; D008024:Ligands; D011485:Protein Binding
D008024:Ligands; D000990:Antithrombin III; D002482:Cellulose; D006493:Heparin; C012024:rayon, purified