Adsorptive membrane chromatography for purification of plasmid DNA.
Adsorptive membranes were investigated for the downstream processing of plasmid DNA by quantifying both separation efficiencies and adsorption uptake with the anion-exchange membranes. Separation efficiencies of the 10-ml Mustang-Q were measured using pulses of 6.1-kilo base pair plasmid DNA and lysozyme tracers, and comparing the responses for both conventional and reverse-flow operation. The plasmid exhibited nearly 200 plates/cm, almost as high efficiency as the protein despite the large difference in size. This behavior contrasts strongly with typical behavior for spherical porous particle packings, which predicted large decreases in efficiency with increases in tracer size. Batch adsorption isotherms for the 6.1-kilo base pair plasmid on small sheets of anion-exchange membranes at various ionic strengths showed high capacities for very large biomolecules. The maximum binding capacity for the membrane unit was calculated as 10 mg plasmid/ml, an order of magnitude greater than typical values reported for porous beads.
Teeters
M A
MA
Department of Chemical Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706-1691, USA.
Conrardy
S E
SE
Thomas
B L
BL
Root
T W
TW
Lightfoot
E N
EN
D000327:Adsorption; D000837:Anion Exchange Resins; D002853:Chromatography, Liquid / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D008567:Membranes, Artificial; D010957:Plasmids
D000837:Anion Exchange Resins; D008567:Membranes, Artificial; D004247:DNA