Purification of plasmid DNA from Escherichia coli ferments using anion-exchange membrane and hydrophobic chromatography

Patricia Guerrero-Germán*, Rosa Ma Montesinos-Cisneros, D. Miguel F. Prazeres, Armando Tejeda-Mansir

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

A novel downstream bioprocess was developed to obtain purified plasmid DNA (pDNA) from Escherichia coli ferments. The intermediate recovery and purification of the pDNA in cell lysate was conducted using hollow-fiber tangential filtration and frontal anion-exchange membrane and elution hydrophobic chromatographies. The purity of the solutions of pDNA obtained during each process stage was investigated. The results show that the pDNA solution purity increased 30-fold and more than 99% of RNA in the lysate was removed during the process operations. The combination of membrane operations and hydrophobic interaction chromatography resulted in an efficient way to recover pDNA from cell lysates. A better understanding of membrane-based technology for the purification of pDNA from clarified E. coli lysate was developed in this research.

Original languageEnglish
Pages (from-to)68-74
Number of pages7
JournalBiotechnology and Applied Biochemistry
Volume58
Issue number1
DOIs
StatePublished - Jan 2011

Keywords

  • Hydrophobic chromatography
  • Ion-exchange chromatography
  • Membrane chromatography
  • Plasmid DNA production
  • RNA removed
  • Tangential filtration

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