BIOT 439 |
| Recombinant plasmid DNA can be used as a biotherapeutic for gene therapy and DNA vaccination. Although various chromatographic methods have been examined, these are expensive and time-consuming for large scale DNA purification due to the low binding capacity. The objective of this work was to evaluate the use of membrane ultrafiltration for plasmid DNA purification including the effects of flux and solution conditions on plasmid transmission. Stirred-cell sieving experiments were conducted with Ultracel membranes using a 3 kbp plasmid. At constant flux, the plasmid transmission increased over 80-fold as the NaCl concentration increased from 0 to 150 mM or as the MgCl2 concentration increased from 0 to 10 mM. Higher MgCl2 concentrations (up to 40 mM) showed no significant differences in the transmission behavior compared to 10 mM. Plasmid transmission was also a strong function of filtrate flux due to DNA elongation. These results provide important insights into the factors governing plasmid ultrafiltration. |
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Downstream Processing: Non-Chromatographic Separation Techniques: Improving Process Throughput
8:00 AM-11:00 AM, Thursday, August 23, 2007 BCEC -- 107B, Oral
Division of Biochemical Technology |