ABSTRACT
Albumin has been purified by chromatography using standardized procedures with different starting materials depending on previous fractionation. Plasma can be used directly after cryoprecipitation and Factor IX adsorption or after isolation of IgG. The plasma was centrifuged and then desalted on Sephadex G-25 Coarse, the pH was adjusted, and the euglobulins were precipitated before ion-exchange chromatography on DEAE- and CM-Sepharose CL-6B. This was followed by concentration by ultrafiltration, gel filtration on Sephacryl S-200, and final concentration and formulation. The albumin obtained was 99% pure and contained less than 1% of aggregated protein.
Subject(s)
Serum Albumin/isolation & purification , Chromatography, Ion Exchange/methods , Electrophoresis, Polyacrylamide Gel , Humans , Immunoelectrophoresis, Two-Dimensional , Immunoglobulin G/isolation & purification , SpectrophotometryABSTRACT
Albumin is obtainable from human blood plasma by an ion exchange chromatographic procedure in a yield of about 95% and a purity well above Pharmacopoeia requirements. Cryosupernatant, factor IX depleted plasma is precipitated with 12 and 25% w/v polyethylene glycol 4000. The second precipitate is dissolved to 8% w/v protein and applied to a DEAE-Sephadex A-50 or a DEAE-Sepharose CL-6B column. Albumin is further purified by chromatography on SP-Sephadex C-50. Gel filtration on Sephadex G-25 is used for desalting prior to lyophilization. The process has been initially designed for fractionation of 50 litres plasma/week but can be further scaled up to meet considerably higher capacity requirements.
