Enhancement of anion-exchange chromatography of DNA using compaction agents

Jason C. Murphy, George E. Fox, Richard C. Willson

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

The use of adsorptive chromatography for preparative nucleic acid separations is often limited by low capacity. The possibility that the adsorbent surface area sterically accessible to nucleic acid molecules could be increased by reducing their radius of gyration with compaction agents has been investigated. The equilibrium adsorption capacity of Q Sepharose anion-exchange matrix for plasmid DNA at 600 mM NaCl was enhanced by up to ca. 40% in the presence of 2.5 mM spermine. In addition, compaction agent selectivity has been demonstrated. Spermine, for example, enhances the adsorption of both plasmid and genomic DNA, spermidine enhances binding only of plasmid, and hexamine cobalt enhances only the binding of genomic DNA. Compaction may be generally useful for enhancing adsorptive separations of nucleic acids.

Original languageEnglish (US)
Pages (from-to)215-221
Number of pages7
JournalJournal of Chromatography A
Volume984
Issue number2
DOIs
StatePublished - Jan 17 2003

Keywords

  • Adsorption
  • Compaction agents
  • DNA
  • Hexamine cobalt
  • Preparative chromatography
  • Spermidine
  • Spermine

ASJC Scopus subject areas

  • Analytical Chemistry

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