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*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Scopus citations


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

Bibliographical note

Funding Information:
Acknowledgments The authors gratefully acknowledge support of this work by the Consejo Nacional de Ciencia y Tecnología de México under grant U39963-Z and fellowship 145061 (P. Guerrero), Department of Chemical and Biological Engineering of the Centre for Biological and Chemical Engineering, Instituto Superior Técnico of the Lisbon University, CINVESTAV-IPN and the Universidad de Sonora. We also gratefully acknowledge the excellent scientific assistance of the PhD student Ana Gabriela Gomes.


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


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