Capturing Enveloped Viruses on Affinity Grids for Downstream Cryo- Electron Microscopy Applications

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dc.contributorVirginia Techen
dc.contributor.authorKiss, G.en
dc.contributor.authorChen, X. M.en
dc.contributor.authorBrindley, M. A.en
dc.contributor.authorCampbell, P.en
dc.contributor.authorAfonso, C. L.en
dc.contributor.authorKe, Z. L.en
dc.contributor.authorHoll, J. M.en
dc.contributor.authorGuerrero-Ferreira, R. C.en
dc.contributor.authorByrd-Leotis, L. A.en
dc.contributor.authorSteel, J.en
dc.contributor.authorSteinhauer, D. A.en
dc.contributor.authorPlemper, R. K.en
dc.contributor.authorKelly, Deborah F.en
dc.contributor.authorSpearman, P. W.en
dc.contributor.authorWright, E. R.en
dc.date.accessed2014-07-15en
dc.date.accessioned2014-07-21T15:49:40Zen
dc.date.available2014-07-21T15:49:40Zen
dc.date.issued2014-02en
dc.description.abstractElectron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.en
dc.description.sponsorshipEmory University, Children's Healthcare of Atlanta, the Center for AIDS Research at Emory University (P30 AI050409)en
dc.description.sponsorshipGeorgia Research Allianceen
dc.description.sponsorshipNSF grant 0923395en
dc.description.sponsorshipNIH/NIAID public health service grants AI101775, AI058828, AI083402en
dc.description.sponsorshipUSDA_ARS CRIS project number 66612-32000-064en
dc.description.sponsorshipCenters for Excellence in Influenza Research and Surveillance (CEIRS) HHSN266200700006Cen
dc.identifier.citationKiss, G.; Chen, X. M.; et al., "Capturing Enveloped Viruses on Affinity Grids for Downstream Cryo- Electron Microscopy Applications," Microsc. Microanal. 20, 164-174, 2014. DOI: 10.1017/s1431927613013937en
dc.identifier.doihttps://doi.org/10.1017/s1431927613013937en
dc.identifier.issn1431-9276en
dc.identifier.urihttp://hdl.handle.net/10919/49642en
dc.identifier.urlhttp://journals.cambridge.org/action/displayFulltext?type=1&fid=9159612&jid=MAM&volumeId=20&issueId=01&aid=9159609&bodyId=&membershipNumber=&societyETOCSession=en
dc.language.isoen_USen
dc.publisherCambridge University Pressen
dc.subjectcryo-electron microscopy (cryo-em)en
dc.subjectcryo-electron tomography (cryo-et)en
dc.subjectaffinity grid (ag)en
dc.subjectenveloped virusen
dc.subjectrespiratory syncytial virusen
dc.subjectinfluenza-a virusen
dc.subjectdefective interferingen
dc.subjectparticlesen
dc.subjectrous-sarcoma-virusen
dc.subjectcryoelectron tomographyen
dc.subjectmonolayeren
dc.subjectpurificationen
dc.subjectmeasles-virusen
dc.subjectvirionsen
dc.subjectcryotomographyen
dc.subjectrevealsen
dc.subjectmaterials science, multidisciplinaryen
dc.subjectmicroscopyen
dc.titleCapturing Enveloped Viruses on Affinity Grids for Downstream Cryo- Electron Microscopy Applicationsen
dc.title.serialMicroscopy and Microanalysisen
dc.typeArticle - Refereeden

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