Dean vortex membrane microfiltration and diafiltration of rBDNF E. coli inclusion bodies.
Cross-flow microfiltration (CMF) and diafiltration were used to concentrate and purify recombinant Brain-Derived Neutrophic Factor (rBDNF) inclusion bodies from an E. coli cell suspension and a homogenized E. coli cell suspension (homogenate/lysate). Although these processes have been tested industrially in pilot scale with conventional linear membrane microfiltration modules, their performances were severely limited due to membrane fouling. The purpose of this work was to determine whether Dean vortex microfiltration with controlled centrifugal instabilities (Dean vortices produced in helical flow) could be used to improve filtration performance over that observed with conventional linear cross-flow microfiltration (CMF). For the microfiltration experiments with the feeds containing cell and homogenate suspensions, improvements in flux of about 50 and 70%, respectively, were obtained with the helical module as compared with that obtained with the linear module. For diafiltration with the homogenate suspension as feed, solute transport (as measured by mass) was from 100 to 40% higher after 40 and 100 min, respectively, with the helical module as compared with that obtained with the linear module. In the presence of the neutral surfactant, Tween 20, solute transport for diafiltration was at least 25 times higher during the first 10 min of operation and 100% higher after 300 min with the helical module as compared with that obtained with the linear module. Clearly, improved filtration performance, a purer and more concentrated product, and substantial savings can be expected with the new Dean vortex filters.
Schutyser
Maarten
M
Howard P. Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, USA.
Rupp
Randall
R
Wideman
Janusz
J
Belfort
Georges
G
D019208:Brain-Derived Neurotrophic Factor / Q000737:chemistry / Q000302:isolation & purification*; D002498:Centrifugation / Q000379:methods; D004867:Equipment Design; D004926:Escherichia coli; D005374:Filtration / Q000295:instrumentation / Q000379:methods*; D002479:Inclusion Bodies / Q000737:chemistry*; D008567:Membranes, Artificial; D008851:Micropore Filters; D008958:Models, Molecular; D010539:Permeability; D011136:Polysorbates / Q000737:chemistry; D011312:Pressure; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012212:Rheology; D012680:Sensitivity and Specificity
D019208:Brain-Derived Neurotrophic Factor; D008567:Membranes, Artificial; D011136:Polysorbates; D011994:Recombinant Proteins