Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA

Clarence M. Ongkudon, and Hodges E, and Murphy K, and Danquah MK, (2012) Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA. Journal of Separation Science, 35 (22). pp. 3208-3216. ISSN 1615-9306

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Abstract

Endotoxin lipopolysaccharide removal from plasmid DNA-based vaccine remains a very challenging task for bioprocess engineers. This paper examined the potential use and advantages of divalent cation (Zn(2+), Ca(2+), Mg(2+)) induced aggregation as a plasmid DNA purification method for lipopolysaccharide removal. Analysis of zeta potential, hydrodynamic size, percentage of aggregation; UV-Vis spectroscopy and electron microscopy were performed to determine the optimal cation for preferential aggregation of lipopolysaccharide over plasmid DNA. The results from the hydrodynamic size analysis showed that the addition of Zn(2+) resulted in the maximum theoretical number of lipopolysaccharide molecules per aggregate particle. Dynamic light scattering analysis showed that plasmid DNA aggregates formed a larger maximum hydrodynamic size when it was treated with Ca(2+) than the other two cations. The K(m) value for lipopolysaccharide-Zn(2+) was substantially low (0.28 M) and considerably large (>2 M) for plasmid DNA-Zn(2+). Scatchard plots for plasmid DNA cations showed positive slopes indicating that there was a minimum concentration of plasmid DNA or cations before a significant aggregation occurred. This work concluded that Zn(2+) had the most preferential aggregative interaction with lipopolysaccharide compared to Mg(2+) and Ca(2+).

Item Type: Article
Uncontrolled Keywords: Endotoxin lipopolysaccharide , vaccine , plasmid DNA , hydrodynamic
Subjects: Q Science > QH Natural history
Divisions: SCHOOL > Biotechnology Research Institute (BRI)
Depositing User: Noraini
Date Deposited: 10 Jul 2018 07:04
Last Modified: 10 Jul 2018 07:04
URI: http://eprints.ums.edu.my/id/eprint/20417

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