Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography

Patricia Guerrero-Germán, Duarte M.F. Prazeres, Roberto Guzmán, Rosa Ma Montesinos-Cisneros, Armando Tejeda-Mansir

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

A new bioprocess using mainly membrane operations to obtain purified plasmid DNA from Escherechia coli ferments was developed. The intermediate recovery and purification of the plasmid DNA in cell lysate was conducted using hollow-fiber tangential filtration and tandem anion-exchange membrane chromatography. The purity of the solutions of plasmid DNA obtained during each process stage was investigated. The results show that more than 97% of RNA in the lysate was removed during the process operations and that the plasmid DNA solution purity increased 28-fold. One of the main characteristics of the developed process is to avoid the use of large quantities of precipitating agents such as salts or alcohols. A better understanding of membrane-based technology for the purification of plasmid DNA from clarified E. coli lysate was developed in this research. The convenience of anion-exchange membranes, configured in ready-to-use devices can further simplify large-scale plasmid purification strategies.

Original languageEnglish (US)
Pages (from-to)615-623
Number of pages9
JournalBioprocess and Biosystems Engineering
Volume32
Issue number5
DOIs
StatePublished - Aug 2009

Keywords

  • Ion-exchange
  • Membrane chromatography
  • Membrane filtration
  • Plasmid DNA

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

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