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dc.contributor.authorOngkudon, C.
dc.contributor.authorDanquah, Michael
dc.identifier.citationOngkudon, C. and Danquah, M. 2010. Process optimisation for anion exchange monolithic chromatography of 4.2kbp plasmid vaccine (pcDNA3F). Journal of Chromatography B. 878 (28): pp. 2719-2725.

Anion exchange monolithic chromatography is increasingly becoming a prominent tool for plasmid DNA purification but no generic protocol is available to purify all types of plasmid DNA. In this work, we established a simple framework and used it to specifically purify a plasmid DNA model from a clarified alkaline-lysed plasmid-containing cell lysate. The framework involved optimising ligand functionalisation temperature (30-80°C), mobile phase flow rate (0.1-1.8 mL/min), monolith pore size (done by changing the porogen content in the polymerisation reaction by 50-80%), buffer pH (6-10), ionic strength of binding buffer (0.3-0.7. M) and buffer gradient elution slope (1-10% buffer B/min). We concluded that preferential pcDNA3F adsorption and optimum resolution could be achieved within the tested conditions by loading the clarified cell lysate into 400. nm pore size of monolith in 0.7. M NaCl (pH 6) of binding buffer followed by increasing the NaCl concentration to 1.0. M at 3%B/min. © 2010 Elsevier B.V.

dc.publisherElsevier BV
dc.titleProcess optimisation for anion exchange monolithic chromatography of 4.2kbp plasmid vaccine (pcDNA3F)
dc.typeJournal Article
dcterms.source.titleJournal of Chromatography B
curtin.departmentCurtin Sarawak
curtin.accessStatusFulltext not available

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