title: Potential application of hydrogel-based strong anion-exchange membrane for plasmid DNA purification.
abstract: The potential application of a hydrogel-based strong anion-exchange (Q) membrane to purify plasmid DNAs was evaluated. The maximum binding capacity of plasmid DNA was estimated to be 12.4 mg/ml of membrane volume with a plasmid recovery yield of ∼90%. The effect of the inherent properties of plasmid DNA, membrane adsorbent, and the ionic environment on membrane performance was systematically investigated. Plasmid DNAs with smaller tertiary structure tended to have a better recovery than those with larger tertiary structure. Environmental Scanning Electron Microscopy (ESEM) revealed that the hydrogel structure is more porous on one side of membrane than the other. Membrane pre-treatment significantly improved pore distribution and increased membrane porosity resulting in a better adsorption, recovery, and higher flux. The selection of proper operating pH led to further improvement. The relative contribution of these factors to improve membrane chromatography of plasmid DNAs was analyzed using statistical modeling. It was found that the adsorption of plasmid DNA was mainly affected by the available adsorptive area associated with membrane porosity, whereas the recovery of plasmid DNAs was mainly affected by the environmental pH.
authors.affiliation: Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, Ontario, Canada N2L 3G1.
authors.forename: Luyang
authors.initials: L
authors.lastname: Zhong
authors.forename: Jeno
authors.initials: J
authors.lastname: Scharer
authors.forename: Murray
authors.initials: M
authors.lastname: Moo-Young
authors.forename: Drew
authors.initials: D
authors.lastname: Fenner
authors.forename: Lisa
authors.initials: L
authors.lastname: Crossley
authors.forename: C Howie
authors.initials: CH
authors.lastname: Honeyman
authors.forename: Shing-Yi
authors.initials: SY
authors.lastname: Suen
authors.forename: C Perry
authors.initials: CP
authors.lastname: Chou
mesh_terms: D000838:Anions / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004278:DNA, Superhelical / Q000737:chemistry / Q000302:isolation & purification*; D004926:Escherichia coli; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008855:Microscopy, Electron, Scanning; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000838:Anions; D004278:DNA, Superhelical; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate