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Virus purification by CsCl density gradient using general centrifugation

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Abstract

Virus purification by cesium chloride (CsCl) density gradient, which generally requires an expensive ultracentrifuge, is an essential technique in virology. Here, we optimized virus purification by CsCl density gradient using general centrifugation (40,000 × g, 2 h, 4 °C), which showed almost the same purification ability as conventional CsCl density gradient ultracentrifugation (100,000 × g, 1 h, 4 °C) using phages S13′ and φEF24C. Moreover, adenovirus strain JM1/1 was also successfully purified by this method. We suggest that general centrifugation can become a less costly alternative to ultracentrifugation for virus purification by CsCl densiy gradient and will thus encourage research in virology.

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References

  1. Aguirre de Carcer D, Lopez-Bueno A, Pearce DA, Alcami A (2015) Biodiversity and distribution of polar freshwater DNA viruses. Sci Adv 1:e1400127. doi:10.1126/sciadv.1400127

    Article  PubMed  PubMed Central  Google Scholar 

  2. Duffy AM, O’Doherty AM, O’Brien T, Strappe PM (2005) Purification of adenovirus and adeno-associated virus: comparison of novel membrane-based technology to conventional techniques. Gene Ther 12(Suppl 1):S62–S72. doi:10.1038/sj.gt.3302616

    Article  CAS  PubMed  Google Scholar 

  3. Fischer MG, Hackl T (2016) Host genome integration and giant virus-induced reactivation of the virophage mavirus. Nature 540:288–291. doi:10.1038/nature20593

    Article  CAS  PubMed  Google Scholar 

  4. Callens N, Brugger B, Bonnafous P, Drobecq H, Gerl MJ, Krey T, Roman-Sosa G, Rumenapf T, Lambert O, Dubuisson J, Rouille Y (2016) Morphology and molecular composition of purified bovine viral diarrhea virus envelope. PLoS Pathog 12:e1005476. doi:10.1371/journal.ppat.1005476

    Article  PubMed  PubMed Central  Google Scholar 

  5. Bourdin G, Schmitt B, Marvin Guy L, Germond JE, Zuber S, Michot L, Reuteler G, Brussow H (2014) Amplification and purification of T4-like Escherichia coli phages for phage therapy: from laboratory to pilot scale. Appl Environ Microbiol 80:1469–1476. doi:10.1128/AEM.03357-13

    Article  PubMed  PubMed Central  Google Scholar 

  6. Bogerd HP, Kennedy EM, Whisnant AW, Cullen BR (2017) Induced packaging of cellular microRNAs into HIV-1 virions can inhibit infectivity. MBio 8:e02125–e02216. doi:10.1128/mBio.02125-16

    Article  PubMed  PubMed Central  Google Scholar 

  7. Tomono T, Hirai Y, Okada H, Adachi K, Ishii A, Shimada T, Onodera M, Tamaoka A, Okada T (2016) Ultracentrifugation-free chromatography-mediated large-scale purification of recombinant adeno-associated virus serotype 1 (rAAV1). Mol Ther Methods Clin Dev 3:15058. doi:10.1038/mtm.2015.58

    Article  PubMed  PubMed Central  Google Scholar 

  8. Bachrach U, Friedmann A (1971) Practical procedures for the purification of bacterial viruses. Appl Microbiol 22:706–715

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Szelechowski M, Bergeron C, Gonzalez-Dunia D, Klonjkowski B (2013) Production and purification of non replicative canine adenovirus type 2 derived vectors. J Vis Exp. doi:10.3791/50833

    PubMed  PubMed Central  Google Scholar 

  10. Brady JN, Winston VD, Consigli RA (1977) Dissociation of polyoma virus by the chelation of calcium ions found associated with purified virions. J Virol 23:717–724

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Kreider JW, Howett MK, Leure-Dupree AE, Zaino RJ, Weber JA (1987) Laboratory production in vivo of infectious human papillomavirus type 11. J Virol 61:590–593

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Takemoto M, Koike M, Mori Y, Yonemoto S, Sasamoto Y, Kondo K, Uchiyama Y, Yamanishi K (2005) Human herpesvirus 6 open reading frame U14 protein and cellular p53 interact with each other and are contained in the virion. J Virol 79:13037–13046. doi:10.1128/JVI.79.20.13037-13046.2005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Farkas T, Sestak K, Wei C, Jiang X (2008) Characterization of a rhesus monkey calicivirus representing a new genus of Caliciviridae. J Virol 82:5408–5416. doi:10.1128/JVI.00070-08

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Furlong DB, Nibert ML, Fields BN (1988) Sigma 1 protein of mammalian reoviruses extends from the surfaces of viral particles. J Virol 62:246–256

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Benfield DA, Nelson E, Collins JE, Harris L, Goyal SM, Robison D, Christianson WT, Morrison RB, Gorcyca D, Chladek D (1992) Characterization of swine infertility and respiratory syndrome (SIRS) virus (isolate ATCC VR-2332). J Vet Diagn Invest 4:127–133. doi:10.1177/104063879200400202

    Article  CAS  PubMed  Google Scholar 

  16. Turgeon N, Toulouse MJ, Martel B, Moineau S, Duchaine C (2014) Comparison of five bacteriophages as models for viral aerosol studies. Appl Environ Microbiol 80:4242–4250. doi:10.1128/aem.00767-14

    Article  PubMed  PubMed Central  Google Scholar 

  17. Calgua B, Carratala A, Guerrero-Latorre L, de Abreu Correa A, Kohn T, Sommer R, Girones R (2014) UVC inactivation of dsDNA and ssRNA viruses in water: UV fluences and a qPCR-based approach to evaluate decay on viral infectivity. Food Environ Virol 6:260–268. doi:10.1007/s12560-014-9157-1

    Article  CAS  PubMed  Google Scholar 

  18. Takemura-Uchiyama I, Uchiyama J, Kato S, Inoue T, Ujihara T, Ohara N, Daibata M, Matsuzaki S (2013) Evaluating efficacy of bacteriophage therapy against Staphylococcus aureus infections using a silkworm larval infection model. FEMS Microbiol Lett 347:52–60. doi:10.1111/1574-6968.12220

    Article  CAS  PubMed  Google Scholar 

  19. Uchiyama J, Rashel M, Maeda Y, Takemura I, Sugihara S, Akechi K, Muraoka A, Wakiguchi H, Matsuzaki S (2008) Isolation and characterization of a novel Enterococcus faecalis bacteriophage phiEF24C as a therapeutic candidate. FEMS Microbiol Lett 278:200–206. doi:10.1111/j.1574-6968.2007.00996.x

    Article  CAS  PubMed  Google Scholar 

  20. Kanda Y (2013) Investigation of the freely available easy-to-use software ‘EZR’ for medical statistics. Bone Marrow Transplant 48:452–458. doi:10.1038/bmt.2012.244

    Article  CAS  PubMed  Google Scholar 

  21. Muroga N, Taharaguchi S, Ohta H, Yamazaki K, Takase K (2006) Pathogenicity of fowl adenovirus isolated from gizzard erosions to immuno-suppressed chickens. J Vet Med Sci 68:289–291

    Article  PubMed  Google Scholar 

  22. Eglon MN, Duffy AM, O’Brien T, Strappe PM (2009) Purification of adenoviral vectors by combined anion exchange and gel filtration chromatography. J Gene Med 11:978–989. doi:10.1002/jgm.1383

    Article  CAS  PubMed  Google Scholar 

  23. Smith AC, Poulin KL, Parks RJ (2009) DNA genome size affects the stability of the adenovirus virion. J Virol 83:2025–2028. doi:10.1128/jvi.01644-08

    Article  CAS  PubMed  Google Scholar 

  24. Li P, Bellett AJ, Parish CR (1984) The structural proteins of chick embryo lethal orphan virus (fowl adenovirus type 1). J Gen Virol 65:1803–1815. doi:10.1099/0022-1317-65-10-1803

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This study was supported by Grant-in-Aid for Young Scientists (15K19095) from the Japan Society for the Promotion of Science.

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Author notes

  1. Tadahiro Nasukawa and Jumpei Uchiyama contributed equally to the work.

Authors and Affiliations

  1. School of Veterinary Medicine, Azabu University, Fuchinobe 1-7-71, Chuo-ku, Sagamihara-shi, Kanagawa, 252-0206, Japan

    Tadahiro Nasukawa, Jumpei Uchiyama, Satoshi Taharaguchi, Sumire Ota, Hironobu Murakami, Keijirou Mizukami & Masahiro Sakaguchi

  2. Kochi University, Kochi, Japan

    Takako Ujihara & Shigenobu Matsuzaki

Authors
  1. Tadahiro Nasukawa
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  2. Jumpei Uchiyama
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  3. Satoshi Taharaguchi
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  4. Sumire Ota
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  5. Takako Ujihara
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  6. Shigenobu Matsuzaki
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  7. Hironobu Murakami
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  8. Keijirou Mizukami
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  9. Masahiro Sakaguchi
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Corresponding author

Correspondence to Jumpei Uchiyama.

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The authors declare that they have no conflict of interest.

Animal handling and research ethics

Animals were handled by veterinarians in strict accordance with good animal practice following the guidelines of Azabu University. This experiment was approved by the Committee for Animal Experimentation at Azabu University (Certificate No. 150318-2).

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Nasukawa, T., Uchiyama, J., Taharaguchi, S. et al. Virus purification by CsCl density gradient using general centrifugation. Arch Virol 162, 3523–3528 (2017). https://doi.org/10.1007/s00705-017-3513-z

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  • DOI: https://doi.org/10.1007/s00705-017-3513-z

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