title: Faster isolation of recombinant factor VIII SQ, with a superporous agarose matrix.
abstract: A superporous agarose matrix was compared with a corresponding homogenous matrix in the isolation of recombinant factor VIII SQ (r-VIII SQ) by immunoaffinity chromatography. As a reference, the commercially available Sepharose FastFlow, used for a similar purification in the industry, was also evaluated. Breakthrough curves are described for flows between 50 and 400 cm/h with pre-purified r-VIII SQ and with cell culture broth. The superporous gel gave the best performance and a 1000-fold purification was obtained in a one-step procedure. The superporous matrix made it possible to increase the throughput about four-fold, presumably due to its better mass transfer properties. The importance of the ligand distribution profile is discussed based upon immunofluorescence microscopy data.
authors.affiliation: Department of Pure and Applied Biochemistry, Lund University, Sweden.
authors.forename: E
authors.initials: E
authors.lastname: Pålsson
authors.forename: A L
authors.initials: AL
authors.lastname: Smeds
authors.forename: A
authors.initials: A
authors.lastname: Petersson
authors.forename: P O
authors.initials: PO
authors.lastname: Larsson
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D017077:Culture Media, Conditioned / Q000737:chemistry; D005169:Factor VIII / Q000302:isolation & purification*; D016650:Fluorescein-5-isothiocyanate; D007158:Immunologic Techniques; D011994:Recombinant Proteins / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012685:Sepharose; D013205:Staphylococcal Protein A
chemical_list: D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D005169:Factor VIII; D012685:Sepharose; D016650:Fluorescein-5-isothiocyanate