title: A highly purified antithrombin III concentrate prepared from human plasma fraction IV-1 by affinity chromatography.
abstract: We describe an improved method for large-scale purification of antithrombin III (AT-III) from human plasma involving heparin affinity chromatography of redissolved fraction IV-1 paste, viral inactivation by heating, followed by a second heparin affinity column. The characteristics of a new heparin affinity resin and the ability to extrapolate process behavior from small-scale (20 ml) to large-scale (40 liter) columns are described. This supports the use of the small-scale column for process optimization and validation studies in compliance with current regulatory requirements for biological products. The process has been characterized by analytical techniques including sodium dodecyl sulfate (SDS), reducing SDS, and nondenaturing polyacrylamide gel electrophoresis; laser desorption time-of-flight mass spectroscopy, and electrospray mass spectroscopy. These results demonstrate that greater than 95% of the protein in the final products is AT-III, which is greater than 95% active as defined by thrombin inhibition.
authors: Plasma Manufacturing Technology, Miles Inc., Clayton, N.C. 27520.
authors: W R
authors: WR
authors: Lebing
authors: D J
authors: DJ
authors: Hammond
authors: J E
authors: JE
authors: Wydick
authors: G A
authors: GA
authors: Baumbach
mesh_terms: D000990:Antithrombin III / Q000302:isolation & purification*; D005591:Chemical Fractionation; D002846:Chromatography, Affinity / Q000379:methods*; D006493:Heparin; D006801:Humans; D009994:Osmolar Concentration
chemical_list: D000990:Antithrombin III; D006493:Heparin