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

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

Research output: Contribution to journalArticle

28 Citations (Scopus)

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

Fingerprint

Chromatography
Purification
Anions
Ion exchange
Plasmids
DNA
Negative ions
Membranes
RNA
Escherichia coli
Alcohols
Salts
Recovery
Fibers
Technology
Equipment and Supplies
Research

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

Cite this

Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography. / Guerrero-Germán, Patricia; Prazeres, Duarte M F; Guzman, Roberto Z; Montesinos-Cisneros, Rosa Ma; Tejeda-Mansir, Armando.

In: Bioprocess and Biosystems Engineering, Vol. 32, No. 5, 08.2009, p. 615-623.

Research output: Contribution to journalArticle

Guerrero-Germán, Patricia ; Prazeres, Duarte M F ; Guzman, Roberto Z ; Montesinos-Cisneros, Rosa Ma ; Tejeda-Mansir, Armando. / Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography. In: Bioprocess and Biosystems Engineering. 2009 ; Vol. 32, No. 5. pp. 615-623.
@article{a676d8a2804540a7926fcecdd393f4cc,
title = "Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography",
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.",
keywords = "Ion-exchange, Membrane chromatography, Membrane filtration, Plasmid DNA",
author = "Patricia Guerrero-Germ{\'a}n and Prazeres, {Duarte M F} and Guzman, {Roberto Z} and Montesinos-Cisneros, {Rosa Ma} and Armando Tejeda-Mansir",
year = "2009",
month = "8",
doi = "10.1007/s00449-008-0284-7",
language = "English (US)",
volume = "32",
pages = "615--623",
journal = "Bioprocess and Biosystems Engineering",
issn = "1615-7591",
publisher = "Springer Verlag",
number = "5",

}

TY - JOUR

T1 - Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography

AU - Guerrero-Germán, Patricia

AU - Prazeres, Duarte M F

AU - Guzman, Roberto Z

AU - Montesinos-Cisneros, Rosa Ma

AU - Tejeda-Mansir, Armando

PY - 2009/8

Y1 - 2009/8

N2 - 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.

AB - 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.

KW - Ion-exchange

KW - Membrane chromatography

KW - Membrane filtration

KW - Plasmid DNA

UR - http://www.scopus.com/inward/record.url?scp=68349139498&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68349139498&partnerID=8YFLogxK

U2 - 10.1007/s00449-008-0284-7

DO - 10.1007/s00449-008-0284-7

M3 - Article

VL - 32

SP - 615

EP - 623

JO - Bioprocess and Biosystems Engineering

JF - Bioprocess and Biosystems Engineering

SN - 1615-7591

IS - 5

ER -