TY - JOUR
T1 - Purification of plasmid DNA from Escherichia coli ferments using anion-exchange membrane and hydrophobic chromatography
AU - Guerrero-Germán, Patricia
AU - Montesinos-Cisneros, Rosa Ma
AU - Prazeres, D. Miguel F.
AU - Tejeda-Mansir, Armando
PY - 2011/1
Y1 - 2011/1
N2 - 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.
AB - 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.
KW - Hydrophobic chromatography
KW - Ion-exchange chromatography
KW - Membrane chromatography
KW - Plasmid DNA production
KW - RNA removed
KW - Tangential filtration
UR - http://www.scopus.com/inward/record.url?scp=79953245624&partnerID=8YFLogxK
U2 - 10.1002/bab.12
DO - 10.1002/bab.12
M3 - Artículo
SN - 0885-4513
VL - 58
SP - 68
EP - 74
JO - Biotechnology and Applied Biochemistry
JF - Biotechnology and Applied Biochemistry
IS - 1
ER -