Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications

Gabriella Kiss, Xuemin Chen, Melinda A. Brindley, Patricia Campbell, Claudio L. Afonso, Zunlong Ke, Jens M. Holl, Ricardo C. Guerrero-Ferreira, Lauren A. Byrd-Leotis, John Steel, David A. Steinhauer, Richard K. Plemper, Deborah F. Kelly, Paul W. Spearman, Elizabeth R. Wright

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Electron 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.

Original languageEnglish (US)
Pages (from-to)164-174
Number of pages11
JournalMicroscopy and Microanalysis
Volume20
Issue number1
DOIs
StatePublished - Feb 2014

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viruses
Viruses
Electron microscopy
affinity
electron microscopy
grids
purification
Purification
lipoproteins
influenza
human immunodeficiency virus
Lipoproteins
Cell culture
destruction
lipids
Lipids
Tomography
tomography
nickel
inclusions

All Science Journal Classification (ASJC) codes

  • Instrumentation

Cite this

Kiss, G., Chen, X., Brindley, M. A., Campbell, P., Afonso, C. L., Ke, Z., ... Wright, E. R. (2014). Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications. Microscopy and Microanalysis, 20(1), 164-174. https://doi.org/10.1017/S1431927613013937
Kiss, Gabriella ; Chen, Xuemin ; Brindley, Melinda A. ; Campbell, Patricia ; Afonso, Claudio L. ; Ke, Zunlong ; Holl, Jens M. ; Guerrero-Ferreira, Ricardo C. ; Byrd-Leotis, Lauren A. ; Steel, John ; Steinhauer, David A. ; Plemper, Richard K. ; Kelly, Deborah F. ; Spearman, Paul W. ; Wright, Elizabeth R. / Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications. In: Microscopy and Microanalysis. 2014 ; Vol. 20, No. 1. pp. 164-174.
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Kiss, G, Chen, X, Brindley, MA, Campbell, P, Afonso, CL, Ke, Z, Holl, JM, Guerrero-Ferreira, RC, Byrd-Leotis, LA, Steel, J, Steinhauer, DA, Plemper, RK, Kelly, DF, Spearman, PW & Wright, ER 2014, 'Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications', Microscopy and Microanalysis, vol. 20, no. 1, pp. 164-174. https://doi.org/10.1017/S1431927613013937

Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications. / Kiss, Gabriella; Chen, Xuemin; Brindley, Melinda A.; Campbell, Patricia; Afonso, Claudio L.; Ke, Zunlong; Holl, Jens M.; Guerrero-Ferreira, Ricardo C.; Byrd-Leotis, Lauren A.; Steel, John; Steinhauer, David A.; Plemper, Richard K.; Kelly, Deborah F.; Spearman, Paul W.; Wright, Elizabeth R.

In: Microscopy and Microanalysis, Vol. 20, No. 1, 02.2014, p. 164-174.

Research output: Contribution to journalArticle

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T1 - Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications

AU - Kiss, Gabriella

AU - Chen, Xuemin

AU - Brindley, Melinda A.

AU - Campbell, Patricia

AU - Afonso, Claudio L.

AU - Ke, Zunlong

AU - Holl, Jens M.

AU - Guerrero-Ferreira, Ricardo C.

AU - Byrd-Leotis, Lauren A.

AU - Steel, John

AU - Steinhauer, David A.

AU - Plemper, Richard K.

AU - Kelly, Deborah F.

AU - Spearman, Paul W.

AU - Wright, Elizabeth R.

PY - 2014/2

Y1 - 2014/2

N2 - Electron 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.

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