title: Plasmid DNA purification using a multimodal chromatography resin.
abstract: Multimodal chromatography is widely used for isolation of proteins because it often results in improved selectivity compared to conventional separation resins. The binding potential and chromatographic behavior of plasmid DNA have here been examined on a Capto Adhere resin. Capto Adhere is a recent multimodal chromatography material allowing molecular recognition between the ligand and target molecule, which is based on combined ionic and aromatic interactions. Capto Adhere proved to offer a very strong binding of nucleic acids. This property could be used to isolate plasmid DNA from a crude Escherichia coli extract. Using a stepwise NaCl gradient, pure plasmid DNA could be obtained without protein and endotoxin contaminations. The RNA fraction bound most strongly to the resin and could be eluted only at very high salt concentrations (2.0 M NaCl). The chromatographic separation behavior was very robust between pH values 6 and 9, and the dynamic binding capacity was estimated to 60 µg/ml resin.
authors.affiliation: Department of Pure and Applied Biochemistry, Lund University, SE 22100, Lund, Sweden; CICS - Health Sciences Research Centre, University of Beira Interior, 6201-001, Covilhã, Portugal.
authors.forename: Tiago
authors.initials: T
authors.lastname: Matos
authors.forename: João A
authors.initials: JA
authors.lastname: Queiroz
authors.forename: Leif
authors.initials: L
authors.lastname: Bülow
mesh_terms: D002845:Chromatography; D004247:DNA / Q000302:isolation & purification*; D004926:Escherichia coli; D008024:Ligands; D010957:Plasmids / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification
chemical_list: D008024:Ligands; D012313:RNA; D004247:DNA
keywords: RNA; endotoxin; hydrophobic interaction; ionic recognition; multimodal chromatography; plasmid DNA