title: Supercoiled plasmid DNA: selective purification by thiophilic/aromatic adsorption.
abstract: Separation of the different plasmid isoforms is a major challenge in purifying plasmid DNA. We describe a new type of biochemical interaction that occurs in the presence of high concentrations of lyotropic salt and results in the selective adsorption of supercoiled plasmid DNA to aromatic thioether ligands. Under well-defined conditions, these ligands are capable of separating supercoiled plasmid DNA (ccc) from its isoform, i.e. open circular (oc) form. Integrated in a process, preceded by group separation and followed by anion-exchange chromatography, this new purification method may facilitate the production of highly purified supercoiled plasmid DNA for use in gene therapy and DNA vaccine applications.
authors.affiliation: Amersham Biosciences AB, Bjorkgatan 30, SE-751 84, Uppsala, Sweden. raf.lemmens@eu.amershambiosciences.com
authors.forename: Raf
authors.initials: R
authors.lastname: Lemmens
authors.forename: Urban
authors.initials: U
authors.lastname: Olsson
authors.forename: Tomas
authors.initials: T
authors.lastname: Nyhammar
authors.forename: Joachim
authors.initials: J
authors.lastname: Stadler
mesh_terms: D000327:Adsorption; D002852:Chromatography, Ion Exchange / Q000379:methods; D004278:DNA, Superhelical / Q000302:isolation & purification*; D010957:Plasmids / Q000302:isolation & purification*
chemical_list: D004278:DNA, Superhelical