Supercoiled plasmid DNA: selective purification by thiophilic/aromatic adsorption.
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.
Lemmens
Raf
R
Amersham Biosciences AB, Bjorkgatan 30, SE-751 84, Uppsala, Sweden. raf.lemmens@eu.amershambiosciences.com
Olsson
Urban
U
Nyhammar
Tomas
T
Stadler
Joachim
J
D000327:Adsorption; D002852:Chromatography, Ion Exchange / Q000379:methods; D004278:DNA, Superhelical / Q000302:isolation & purification*; D010957:Plasmids / Q000302:isolation & purification*
D004278:DNA, Superhelical