Purification of plasmid DNA using tangential flow filtration and tandem anion-exchange membrane chromatography. 2009

Patricia Guerrero-Germán, and Duarte M F Prazeres, and Roberto Guzmán, and Rosa Ma Montesinos-Cisneros, and Armando Tejeda-Mansir
Instituto de Ingeniería, Universidad Autónoma de Baja California, Blvd. Benito Juárez y Calle Normal, Col. Insurgentes Este, 21280, Mexicali, Baja California, Mexico.

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.

UI MeSH Term Description Entries
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D002498 Centrifugation Process of using a rotating machine to generate centrifugal force to separate substances of different densities, remove moisture, or simulate gravitational effects. It employs a large motor-driven apparatus with a long arm, at the end of which human and animal subjects, biological specimens, or equipment can be revolved and rotated at various speeds to study gravitational effects. (From Websters, 10th ed; McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
D002852 Chromatography, Ion Exchange Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins. Chromatography, Ion-Exchange,Ion-Exchange Chromatography,Chromatographies, Ion Exchange,Chromatographies, Ion-Exchange,Ion Exchange Chromatographies,Ion Exchange Chromatography,Ion-Exchange Chromatographies
D000837 Anion Exchange Resins High-molecular-weight insoluble polymers that contain functional cationic groups capable of undergoing exchange reactions with anions. Anion Exchange Resin,Anion Exchangers (Resins),Exchange Resin, Anion,Exchange Resins, Anion,Resin, Anion Exchange,Resins, Anion Exchange
D014462 Ultrafiltration The separation of particles from a suspension by passage through a filter with very fine pores. In ultrafiltration the separation is accomplished by convective transport; in DIALYSIS separation relies instead upon differential diffusion. Ultrafiltration occurs naturally and is a laboratory procedure. Artificial ultrafiltration of the blood is referred to as HEMOFILTRATION or HEMODIAFILTRATION (if combined with HEMODIALYSIS).
D044085 Microfluidics The study of fluid channels and chambers of tiny dimensions of tens to hundreds of micrometers and volumes of nanoliters or picoliters. This is of interest in biological MICROCIRCULATION and used in MICROCHEMISTRY and INVESTIGATIVE TECHNIQUES. Microfluidic

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