1. Plasmid DNA pre-purification by tangential flow filtration
- Author
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Rosa Maria Montesinos-Cisneros, Patricia Guerrero-Germán, Ivan Manzano, and Armando Tejeda-Mansir
- Subjects
Chromatography ,Chemistry ,Ultrafiltration ,medicine.disease_cause ,Molecular biology ,Membrane technology ,Cross-flow filtration ,Diafiltration ,Membrane ,Plasmid ,Plasmid dna ,medicine ,Escherichia coli ,Biotechnology - Abstract
The growing demand of plasmid DNA for gene therapy and third generation vaccines has encouraged the development of simple, robust and scalable production bioprocesses. Bearing in mind the purification step, the membrane separation processes may be a viable and cost-effective option that may be used without compromising the plasmid integrity. In this work, an operation of tangential flow ultrafiltration in a hollow-fiber module was used for the pre-purification of plasmid pVAX1-NH36 from clarified lysates of Escherichia coli DH5α culture. The ultrafiltration system was characterized using model solutions to determine the membrane resistance, plasmid concentration on the membrane wall and the system mass-transfer coefficient. In the analysis, a model based on Darcy's law and the stagnant film layer was used. These results allowed to estimate an optimal flux of 0.0016 cm/s and an optimal bulk concentration for the diafiltration process of 5.50 µg/mL. The concentration of lysates containing the plasmid of int...
- Published
- 2015