Flocculation: an alternative process to ion-exchange chromatography: (A scale-up study using recombinant human superoxide dismutase as model protein).
Flocculation using pellicular charged flocculants was investigated as an alternative process to conventional chromatography in purification of recombinant proteins using human recombinant superoxide dismutase expressed in E. coli as a model system. The removal of pyrogens, proteins, debris and the yield were determined. At laboratory scale, the starting conditions were optimized to yield a stable solution and the flocculation process fitted into a purification scheme. 100 L fermentation broth was the initial volume at pilot scale. The process parameters determined at the laboratory scale were tested and the results were compared to the pilot scale. The method was also compared to an ion-exchange chromatography.
Institute of Applied Microbiology, University of Bodenkultur Nussdorferlände 11, Vienna, Austria.
K
K
Vorauer-Uhl
M
M
Skias
A
A
Trkola
W
W
Schönhofer
F
F
Unterluggauer
P
P
Schulz
A
A
Jungbauer
D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005285:Fermentation; D005427:Flocculation; D006801:Humans; D008956:Models, Chemical; D010865:Pilot Projects; D011994:Recombinant Proteins / Q000302:isolation & purification*; D013482:Superoxide Dismutase / Q000235:genetics / Q000302:isolation & purification*
D011994:Recombinant Proteins; D013482:Superoxide Dismutase