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Stability, biophysical properties and effect of ultracentrifugation and diafiltration on measles virus and mumps virus

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Abstract

Measles virus and mumps virus (MeV and MuV) are enveloped RNA viruses used for production of live attenuated vaccines for prophylaxis of measles and mumps disease, respectively. For biotechnological production of and basic research on these viruses, the preparation of highly purified and infectious viruses is a prerequisite, and to meet that aim, knowledge of their stability and biophysical properties is crucial. Our goal was to carry out a detailed investigation of the stability of MeV and MuV under various pH, temperature, shear stress, filtration and storage conditions, as well as to evaluate two commonly used purification techniques, ultracentrifugation and diafiltration, with regard to their efficiency and effect on virus properties. Virus titers were estimated by CCID50 assay, particle size and concentration were measured by Nanoparticle tracking analysis (NTA) measurements, and the host cell protein content was determined by ELISA. The results demonstrated the stability of MuV and MeV at pH <9 and above pH 4 and 5, respectively, and aggregation was observed at pH >9. Storage without stabilizer did not result in structural changes, but the reduction in infectivity after 24 hours was significant at +37 °C. Vortexing of the viruses resulted in significant particle degradation, leading to lower virus titers, whereas pipetting had much less impact on virus viability. Diafiltration resulted in higher recovery of both total and infectious virus particles than ultracentrifugation. These results provide important data for research on all upstream and downstream processes on these two viruses regarding biotechnological production and basic research.

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Acknowledgements

The authors would like to thank Mrs. Renata Jug for her assistance in the experimental procedures. This study was funded by the Croatian Science Foundation (grant number 8193 to MB).

Any opinions, findings and conclusions or recommendations expressed in this article are those of the authors and do not necessarily reflect the views of the Croatian Science Foundation.

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Authors and Affiliations

  1. Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, HR-10 000, Zagreb, Croatia

    Dora Sviben, Dubravko Forčić, Tihana Kurtović, Beata Halassy & Marija Brgles

  2. Centre of Excellence for Viral Immunology and Vaccines, CERVirVac, Zagreb, Croatia

    Dora Sviben, Dubravko Forčić, Tihana Kurtović, Beata Halassy & Marija Brgles

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  1. Dora Sviben
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  2. Dubravko Forčić
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Correspondence to Dora Sviben.

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Sviben, D., Forčić, D., Kurtović, T. et al. Stability, biophysical properties and effect of ultracentrifugation and diafiltration on measles virus and mumps virus. Arch Virol 161, 1455–1467 (2016). https://doi.org/10.1007/s00705-016-2801-3

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  • DOI: https://doi.org/10.1007/s00705-016-2801-3

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