Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography
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 chroma...
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| Veröffentlicht in: | Bioprocess and biosystems engineering 2009-08, Vol.32 (5), p.615-623 |
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| creator | Guerrero-Germán, Patricia Prazeres, Duarte M. F Guzmán, Roberto Montesinos-Cisneros, Rosa Ma Tejeda-Mansir, Armando |
| description | 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. |
| doi_str_mv | 10.1007/s00449-008-0284-7 |
| format | Article |
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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. 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| identifier | ISSN: 1615-7591 |
| ispartof | Bioprocess and biosystems engineering, 2009-08, Vol.32 (5), p.615-623 |
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| source | MEDLINE; SpringerNature Journals |
| subjects | Anion Exchange Resins - chemistry Biological and medical sciences Biotechnology Centrifugation - methods Chemistry Chemistry and Materials Science Chromatography Chromatography, Ion Exchange - methods Deoxyribonucleic acid DNA E coli Environmental Engineering/Biotechnology Escherichia coli Fermentation Filtration Food Science Fundamental and applied biological sciences. Psychology Industrial and Production Engineering Industrial Chemistry/Chemical Engineering Membranes Microfluidics - methods Original Paper Plasmids - isolation & purification Proteins Studies Ultrafiltration - methods |
| title | Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography |
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