Purification of plasmid DNA with a new type of anion-exchange beads having a non-charged surface

Journal of Chromatography. a
Per-Erik GustavssonPer-Olof Larsson

Abstract

We have prepared a new type of anion exchanger, which effectively discriminates between RNA and plasmid DNA. The material is based on a Sephacryl S-500 HR matrix provided with quartenary amine anion-exchange groups. A distinguishing feature of the beads is that a thin (2-3 microm) outer layer of the beads lacks ion-exchange groups. In the synthesis of these beads the vinyl groups in the outer layer of vinylalkyl substituted Sephacryl S-500 HR beads are reacted with bromine. The resulting layer of bromoalkyl groups are hydrolysed, creating an inert outer layer of hydroxyalkyl groups. Finally, bromination and trimethylamine reactions of the inner vinyl groups provide the beads with a core of cationic groups. Large plasmid molecules will not bind to such beads since they are too large to enter the pores and therefore cannot come into contact with the charged matrix in the inner parts of the beads. RNA and protein molecules present in a cleared lysate, on the other hand, readily enter the pores and become adsorbed. A two-column strategy was developed for plasmid purification (recombinant pBluescript, 5.9 kilo base pairs, kbp). The first column was packed with the restricted access anion-exchanger beads (lid beads) and the second co...Continue Reading

References

Jun 26, 1998·Journal of Chromatography. a·D M PrazeresC Cooney
Sep 22, 1998·Nucleic Acids Research·T Schluep, C L Cooney
Dec 31, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Raf LemmensJoachim Stadler

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Citations

Jun 14, 2006·Bioprocess and Biosystems Engineering·Rosa Ma Montesinos-CisnerosArmando Tejeda-Mansir
May 12, 2009·Journal of Biotechnology·Christina ParilAlois Jungbauer
Aug 4, 2007·Journal of Biotechnology·Nadine StreitnerErwin Flaschel
Apr 4, 2015·Journal of Chromatography. a·Micky Fu Xiang LeeBeng Ti Tey
Jan 1, 2010·Biotechnology & Genetic Engineering Reviews·F SousaJ A Queiroz
Oct 31, 2006·Journal of Chromatography. a·Peter TiainenPer-Olof Larsson
May 8, 2008·Journal of Separation Science·Christina Tarmann, Alois Jungbauer
Sep 11, 2012·Journal of Separation Science·Angela SousaJoão A Queiroz
Jun 19, 2021·Journal of Chromatography. a·Xue MiGiorgio Carta
Mar 11, 2015·ACS Applied Materials & Interfaces·Cláudia S M FernandesA Cecília A Roque
May 31, 2011·Journal of Chromatography. a·Ganesh IyerKwok-Shun Cheng

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