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

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

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 lipopolysacchari...

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Main Authors: Clarence M. Ongkudon, Hodges E, Murphy K, Danquah MK
Format: Article
Language:English
Published: John Wiley & Sons Ltd. 2012
Subjects:
QH Natural history
Online Access:http://eprints.ums.edu.my/20417/
http://eprints.ums.edu.my/20417/
http://eprints.ums.edu.my/20417/1/Removal%20of%20endotoxins%20from%20plasmid%20DNA.pdf
id ums-20417
recordtype eprints
spelling ums-204172018-07-10T07:04:24Z http://eprints.ums.edu.my/20417/ Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA Clarence M. Ongkudon, Hodges E, Murphy K, Danquah MK, QH Natural history 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+). John Wiley & Sons Ltd. 2012 Article PeerReviewed text en http://eprints.ums.edu.my/20417/1/Removal%20of%20endotoxins%20from%20plasmid%20DNA.pdf 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 https://doi.org/10.1002/jssc.201200237
repository_type Digital Repository
institution_category Local University
institution Universiti Sabah Malaysia
building UMS Institutional Repository
collection Online Access
language English
topic QH Natural history
spellingShingle QH Natural history
Clarence M. Ongkudon,
Hodges E,
Murphy K,
Danquah MK,
Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA
description 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+).
format Article
author Clarence M. Ongkudon,
Hodges E,
Murphy K,
Danquah MK,
author_facet Clarence M. Ongkudon,
Hodges E,
Murphy K,
Danquah MK,
author_sort Clarence M. Ongkudon,
title Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA
title_short Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA
title_full Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA
title_fullStr Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA
title_full_unstemmed Removal of endotoxins from plasmid DNA: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid DNA
title_sort removal of endotoxins from plasmid dna: analysis of aggregative interaction of mobile divalent metal cations with endotoxins and plasmid dna
publisher John Wiley & Sons Ltd.
publishDate 2012
url http://eprints.ums.edu.my/20417/
http://eprints.ums.edu.my/20417/
http://eprints.ums.edu.my/20417/1/Removal%20of%20endotoxins%20from%20plasmid%20DNA.pdf
first_indexed 2018-09-05T09:43:20Z
last_indexed 2018-09-05T09:43:20Z
_version_ 1610760166039355392

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