Milligram scale parallel purification of plasmid DNA using anion-exchange membrane capsules and a multi-channel peristaltic pump |
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| Authors: | Syrén Per-Olof Rozkov Aleksei Schmidt Stefan R Strömberg Patrik |
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| Institution: | Global Protein Science and Supply, AstraZeneca R&D S?dert?lje, SE-15185 S?dert?lje, Sweden. |
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| Abstract: | A parallel chromatographic procedure for the purification of milligram amounts of plasmid DNA was developed. Initial studies showed that ion-exchange membrane capsules displayed high capacity for plasmid DNA. Interestingly, a weak anion exchanger (DEAE) proved to be superior to the strong quarternary ammonium group with respect to elution and regeneration properties and the 75 cm(2) Sartobind D membrane capsule (MA75D, Sartorius) was selected for further studies. A method for reducing endotoxin levels by using CTAB as a precipitant was optimised. By introducing this step into the protocol, endotoxin levels could be reduced approximately 100-fold to
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