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Braz. j. med. biol. res ; 29(2): 185-91, Feb. 1996. graf
Article in English | LILACS | ID: lil-161668

ABSTRACT

We have modified a standard column chromatography method for the preparation of albumin from human plasma. The proposed method utilizes Sephadex G-25, euglobulin precipitation, DEAE-Sepharose, ethanol heat treatment and Sephacryl S-200 HR. The procedure differs from that normally used by the introduction of Schneider's ethanol/heat treatment step and the elimination of a CM-Sepharose step, after the DEAE-Sepharose step. The proposed method was used to produce 15 batches of albumin, 100 liters of plasma per batch. Purity of more than 99 percent was obtained at an average yield of 26.5 g/l plasma for the albumin produced. All batches presented 99.2 to 99.9 percent monomer and 0.1 to 0.8 percent dimer, with no larger polymers detected. The utilization of the final gel filtration step eliminated highly polymerized albumin usually obtained during the process. The advantages of the proposed method are reduction in the overall process time from 6 to 5 days, elimination of the CM-Sepharose step and the reduction of 6 to 4 columns in series for the Sephacryl S-200 HR step.


Subject(s)
Humans , Serum Albumin/biosynthesis , Chromatography, Agarose/standards , In Vitro Techniques , Chemical Fractionation , Chromatography/instrumentation
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