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Purification of plasmid DNA from Escherichia coli ferments using anion‐exchange membrane and hydrophobic chromatography
Author(s) -
GuerreroGermán Patricia,
MontesinosCisneros Rosa Ma.,
Prazeres D. Miguel F.,
TejedaMansir Armando
Publication year - 2011
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.12
Subject(s) - lysis , escherichia coli , chromatography , membrane , bioprocess , chemistry , elution , plasmid , ion exchange , ion chromatography , dna , hydrophilic interaction chromatography , biochemistry , biology , high performance liquid chromatography , gene , ion , organic chemistry , paleontology
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.