Single step purification of plasmid DNA using peptide ligand affinity chromatography.
Single step affinity chromatography was employed for the purification of plasmid DNA (pDNA), thus eliminating several steps compared with current commercial purification methods for pDNA. Significant reduction in pDNA production time and cost was obtained. This chromatographic operation employed a peptide-monolith construct to isolate pDNA from Escherichia coli (E. coli) impurities present in a clarified lysate feedstock. Mild conditions were applied to avoid any degradation of pDNA. The effect of some important parameters on pDNA yield was also evaluated with the aim of optimising the affinity purification of pDNA. The results demonstrate that 81% of pDNA was recovered and contaminating gDNA, RNA and protein were removed below detectable levels.
Han
Ying
Y
Bio Engineering Laboratory, Department of Chemical Engineering, Monash University, Wellington Road, Clayton, Melbourne, Vic. 3800, Australia. ying.han@eng.monash.edu.au
Forde
Gareth M
GM
D002846:Chromatography, Affinity / Q000379:methods*; D008024:Ligands; D010455:Peptides / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results
D008024:Ligands; D010455:Peptides