Purification of Viruses and Macromolecular Assemblies for Structural Investigations Using a Novel Ion Exchange Method
Identifieur interne : 004405 ( Main/Exploration ); précédent : 004404; suivant : 004406Purification of Viruses and Macromolecular Assemblies for Structural Investigations Using a Novel Ion Exchange Method
Auteurs : Laura Walin [États-Unis] ; Roman Tuma [États-Unis] ; George J. Thomas Jr. [États-Unis] ; Dennis H. Bamford [États-Unis]Source :
- Virology [ 0042-6822 ] ; 1994.
Abstract
Abstract: We describe a novel ion exchange chromatographic technique suitable for large-scale preparation of viruses and other biomacromolecular assemblies in highly purified form. The method, which utilizes anion exchange on either of two commercially available cellulose cartridges, is applied to the Escherichia coli bacteriophage PRD1. Viral particles eluted from both QMA and DEAE cartridges retain infectivity and exhibit greater homogeneity of composition, as judged by gel electrophoresis and electron microscopy, than particles purified by rate zonal sucrose gradient centrifugation. The ion exchange protocols are rapid, requiring less than 15 min elution time, and permit retrieval of the purified viral particles at high concentration in aqueous media without centrifugal pelleting. The present method is particularly well suited to the preparation of milligram to decigram quantities of virus, sufficient for many biophysical structural analyses, including investigations by solution spectroscopic and crystal diffraction techniques. The feasibility and advantages of the ion exchange chromatographic procedure are demonstrated by application of laser Raman spectroscopy to ion exchange purified PRD1 virions and subviral assemblies.
Url:
DOI: 10.1006/viro.1994.1259
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001695
- to stream Istex, to step Curation: 001695
- to stream Istex, to step Checkpoint: 001A42
- to stream Main, to step Merge: 004468
- to stream Main, to step Curation: 004405
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Purification of Viruses and Macromolecular Assemblies for Structural Investigations Using a Novel Ion Exchange Method</title><author><name sortKey="Walin, Laura" sort="Walin, Laura" uniqKey="Walin L" first="Laura" last="Walin">Laura Walin</name></author><author><name sortKey="Tuma, Roman" sort="Tuma, Roman" uniqKey="Tuma R" first="Roman" last="Tuma">Roman Tuma</name></author><author><name sortKey="Thomas Jr, George J" sort="Thomas Jr, George J" uniqKey="Thomas Jr G" first="George J." last="Thomas Jr.">George J. Thomas Jr.</name></author><author><name sortKey="Bamford, Dennis H" sort="Bamford, Dennis H" uniqKey="Bamford D" first="Dennis H." last="Bamford">Dennis H. Bamford</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:83E31CFD663B681B2AAEC1DA34B41219A201EC53</idno><date when="1994" year="1994">1994</date><idno type="doi">10.1006/viro.1994.1259</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-04KLZ5GP-W/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001695</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001695</idno><idno type="wicri:Area/Istex/Curation">001695</idno><idno type="wicri:Area/Istex/Checkpoint">001A42</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001A42</idno><idno type="wicri:doubleKey">0042-6822:1994:Walin L:purification:of:viruses</idno><idno type="wicri:Area/Main/Merge">004468</idno><idno type="wicri:Area/Main/Curation">004405</idno><idno type="wicri:Area/Main/Exploration">004405</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Purification of Viruses and Macromolecular Assemblies for Structural Investigations Using a Novel Ion Exchange Method</title><author><name sortKey="Walin, Laura" sort="Walin, Laura" uniqKey="Walin L" first="Laura" last="Walin">Laura Walin</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Missouri (État)</region></placeName><wicri:cityArea>Department of Genetics, Institute of Biotechnology, P.O., Box 17, SF-00014 University of Helsinki, Finland; and Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri—Kansas City, Kansas City</wicri:cityArea></affiliation></author><author><name sortKey="Tuma, Roman" sort="Tuma, Roman" uniqKey="Tuma R" first="Roman" last="Tuma">Roman Tuma</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Missouri (État)</region></placeName><wicri:cityArea>Department of Genetics, Institute of Biotechnology, P.O., Box 17, SF-00014 University of Helsinki, Finland; and Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri—Kansas City, Kansas City</wicri:cityArea></affiliation></author><author><name sortKey="Thomas Jr, George J" sort="Thomas Jr, George J" uniqKey="Thomas Jr G" first="George J." last="Thomas Jr.">George J. Thomas Jr.</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Missouri (État)</region></placeName><wicri:cityArea>Department of Genetics, Institute of Biotechnology, P.O., Box 17, SF-00014 University of Helsinki, Finland; and Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri—Kansas City, Kansas City</wicri:cityArea></affiliation></author><author><name sortKey="Bamford, Dennis H" sort="Bamford, Dennis H" uniqKey="Bamford D" first="Dennis H." last="Bamford">Dennis H. Bamford</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Missouri (État)</region></placeName><wicri:cityArea>Department of Genetics, Institute of Biotechnology, P.O., Box 17, SF-00014 University of Helsinki, Finland; and Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri—Kansas City, Kansas City</wicri:cityArea></affiliation></author></analytic><monogr></monogr><series><title level="j">Virology</title><title level="j" type="abbrev">YVIRO</title><idno type="ISSN">0042-6822</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1994">1994</date><biblScope unit="volume">201</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="1">1</biblScope><biblScope unit="page" to="7">7</biblScope></imprint><idno type="ISSN">0042-6822</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0042-6822</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: We describe a novel ion exchange chromatographic technique suitable for large-scale preparation of viruses and other biomacromolecular assemblies in highly purified form. The method, which utilizes anion exchange on either of two commercially available cellulose cartridges, is applied to the Escherichia coli bacteriophage PRD1. Viral particles eluted from both QMA and DEAE cartridges retain infectivity and exhibit greater homogeneity of composition, as judged by gel electrophoresis and electron microscopy, than particles purified by rate zonal sucrose gradient centrifugation. The ion exchange protocols are rapid, requiring less than 15 min elution time, and permit retrieval of the purified viral particles at high concentration in aqueous media without centrifugal pelleting. The present method is particularly well suited to the preparation of milligram to decigram quantities of virus, sufficient for many biophysical structural analyses, including investigations by solution spectroscopic and crystal diffraction techniques. The feasibility and advantages of the ion exchange chromatographic procedure are demonstrated by application of laser Raman spectroscopy to ion exchange purified PRD1 virions and subviral assemblies.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Missouri (État)</li></region></list><tree><country name="États-Unis"><region name="Missouri (État)"><name sortKey="Walin, Laura" sort="Walin, Laura" uniqKey="Walin L" first="Laura" last="Walin">Laura Walin</name></region><name sortKey="Bamford, Dennis H" sort="Bamford, Dennis H" uniqKey="Bamford D" first="Dennis H." last="Bamford">Dennis H. Bamford</name><name sortKey="Thomas Jr, George J" sort="Thomas Jr, George J" uniqKey="Thomas Jr G" first="George J." last="Thomas Jr.">George J. Thomas Jr.</name><name sortKey="Tuma, Roman" sort="Tuma, Roman" uniqKey="Tuma R" first="Roman" last="Tuma">Roman Tuma</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 004405 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 004405 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:83E31CFD663B681B2AAEC1DA34B41219A201EC53
|texte= Purification of Viruses and Macromolecular Assemblies for Structural Investigations Using a Novel Ion Exchange Method
}}
| This area was generated with Dilib version V0.6.33. |  |