Abstract
The analysis of herpes simplex virus type 1 mature extracellular virions by proteomics requires highly enriched samples to limit false positives and favor the detection of true components. The protocol described below involves the removal of highly contaminating serum proteins and purification of the virions by a series of differential and density centrifugation steps. In addition, L-particles, which are viral particles devoid of genome and capsid but present in the extracellular milieu, are depleted on Ficoll 400 gradients. As previously reported, the resulting viral particles are free of most contaminants and suitable for mass spectrometry.
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Acknowledgement
This work was supported by grants from the Canadian Institutes of Health Research (MOP 82921 and MOP 258030). Special thanks to Kerstin Radtke and Nabil El Bilali for proofreading and Christina Bell for the mass spectrometry protocols.
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Lippé, R. (2014). Characterization of Extracellular HSV-1 Virions by Proteomics. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus. Methods in Molecular Biology, vol 1144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0428-0_12
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DOI: https://doi.org/10.1007/978-1-4939-0428-0_12
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