| Differential interactions of plasmid DNA, RNA and genomic DNA with amino acid-based affinity matrices. | |
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MedLine Citation:
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PMID: 20845376 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The development of a strategy to plasmid DNA (pDNA) purification has become necessary for the development of gene therapy and DNA vaccine production processes in recent years, since this nucleic acid and most of contaminants, such as RNA, genomic DNA and endotoxins, are negatively charged. An ideal separation methodology may be achieved with the use of affinity interactions between immobilized amino acids and nucleic acids. In this study, the binding behaviour of nucleic acids under the influence of different environmental conditions, such as the composition and ionic strength of elution buffer, and the temperature, is compared with various amino acids immobilized on chromatography resins. Supercoiled (sc) plasmid isoform was isolated with all matrices used, but in some cases preferential interactions with other nucleic acids were found. Particularly, lysine chromatography showed to be an ideal technology mainly on RNA purification using low salt concentration. On the other hand, arginine ligands have shown a greater ability to retain the sc isoform comparatively to the other nucleic acids retention, becoming this support more adequate to sc pDNA purification. The temperature variation, competitive elution and oligonucleotides affinity studies also allowed to recognize the dominant interactions inherent to biorecognition of pDNA molecule and the affinity matrices. |
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Authors:
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Angela Sousa; Fani Sousa; João A Queiroz |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of separation science Volume: 33 ISSN: 1615-9314 ISO Abbreviation: J Sep Sci Publication Date: 2010 Sep |
Date Detail:
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Created Date: 2010-09-16 Completed Date: 2011-01-13 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101088554 Medline TA: J Sep Sci Country: Germany |
Other Details:
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Languages: eng Pagination: 2610-8 Citation Subset: IM |
Affiliation:
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Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Covilhã, Portugal. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acids
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chemistry* Buffers Chromatography, Affinity / instrumentation*, methods DNA / chemistry* DNA, Superhelical / isolation & purification* Genome* Osmolar Concentration Plasmids / genetics* RNA / chemistry* |
| Chemical | |
Reg. No./Substance:
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0/Amino Acids; 0/Buffers; 0/DNA, Superhelical; 63231-63-0/RNA; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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