title: [Separation of coagulation factor VIII with high activity using gigaporous anion exchange chromatography].
abstract: A purification process to obtain coagulation factor VIII (F VIII) with high activity from human plasma was established. Based on the analysis of the size ratio between F VIII and matrix porous medium and its effect on the protein activity, a novel purification process designed was superporous ion exchange chromatography (IEC). The operating conditions of gigaporous and traditional anion exchange chromatography were optimized separately. The chromogenic substrate, gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to monitor the bioactivity and purity of the chromatographic products. The results showed that the superporous medium could not only protect structure of macro-protein but also enhance its mass transfer, finally giving FVIII product with high activity. The yield of F VIII in superporous chromatography was about five times of commercially agarose chromatography and the specific activity was up to 154 IU/mg protein. Furthermore, we studied the regeneration process of the superporous medium, washing the column with 5 column volumes of 1 mol/L NaOH at a low flow rate, to ensure the chromatographic stability. This purification process is simple, reproducible and suitable for large-scale production.
authors.affiliation: West China School of Pharmacy, Sichuan University, Chengdu 610041, China.
authors.forename: Limei
authors.initials: L
authors.lastname: Kang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Luo
authors.forename: You
authors.initials: Y
authors.lastname: Li
authors.forename: Yuefang
authors.initials: Y
authors.lastname: Zhou
authors.forename: Rong
authors.initials: R
authors.lastname: Yu
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
mesh_terms: D000837:Anion Exchange Resins / Q000737:chemistry*; D000838:Anions / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005169:Factor VIII / Q000302:isolation & purification*; D006801:Humans; D010949:Plasma / Q000737:chemistry*; D016062:Porosity
chemical_list: D000837:Anion Exchange Resins; D000838:Anions; D005169:Factor VIII