Faster isolation of recombinant factor VIII SQ, with a superporous agarose matrix.
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.
Pålsson
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E
Department of Pure and Applied Biochemistry, Lund University, Sweden.
Smeds
A L
AL
Petersson
A
A
Larsson
P O
PO
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
D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D005169:Factor VIII; D012685:Sepharose; D016650:Fluorescein-5-isothiocyanate