Linked Life Data

19635618

Source:http://linkedlifedata.com/resource/pubmed/id/19635618

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
35
pubmed:dateCreated
2009-8-7
pubmed:abstractText
There is considerable interest in using size exclusion chromatography (SEC) to analyze and purify specific plasmid isoforms, but there is currently no fundamental understanding of the effects of plasmid size and morphology on plasmid behavior in SEC. Experiments were performed for plasmids from 3.0 to 17.0kbp in size. The linear and open-circular isoforms were generated from the supercoiled plasmid by appropriate enzymatic digestion. SEC retention data were obtained using a Sephacryl S-1000 SF resin packed column and an Agilent HPLC system over a range of flow rates using buffers of different ionic strength and composition. The plasmid partition coefficients, K(P), were evaluated from the first statistical moment of the chromatographic peak. The partition coefficient decreased with increasing plasmid size as expected; K(P) varied from 0.299 to 0.045 for supercoiled plasmids of 3.0 to 17.0kbp. The partition coefficient also increased with increasing ionic strength due to the compaction of the DNA associated with the shielding of the intramolecular electrostatic interactions. For any plasmid size, the supercoiled isoform had the highest K(P) followed by the open-circular and then the linear isoform, consistent with independent estimates of the plasmid radius of gyration as determined by static light scattering. The experimental data were analyzed using available theoretical models for the partitioning of linear and cyclic polymer chains in well-defined pore geometries. These results provide important insights into the behavior of different plasmid isoforms in size exclusion chromatography.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1873-3778
pubmed:author
pubmed:issnType
Electronic
pubmed:day
28
pubmed:volume
1216
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
6295-302
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Size exclusion chromatography of plasmid DNA isoforms.
pubmed:affiliation
Department of Chemical Engineering, The Pennsylvania State University, University Park, PA 16802, United States.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S.