title: Stability, biophysical properties and effect of ultracentrifugation and diafiltration on measles virus and mumps virus.
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.
authors.affiliation: Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, HR-10 000, Zagreb, Croatia. dsviben@unizg.hr.|Centre of Excellence for Viral Immunology and Vaccines, CERVirVac, Zagreb, Croatia. dsviben@unizg.hr.
authors.forename: Dora
authors.identifier: http://orcid.org/0000-0001-5905-0692
authors.initials: D
authors.lastname: Sviben
authors.affiliation: Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, HR-10 000, Zagreb, Croatia.|Centre of Excellence for Viral Immunology and Vaccines, CERVirVac, Zagreb, Croatia.
authors.forename: Dubravko
authors.initials: D
authors.lastname: Forčić
authors.affiliation: Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, HR-10 000, Zagreb, Croatia.|Centre of Excellence for Viral Immunology and Vaccines, CERVirVac, Zagreb, Croatia.
authors.forename: Tihana
authors.initials: T
authors.lastname: Kurtović
authors.affiliation: Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, HR-10 000, Zagreb, Croatia.|Centre of Excellence for Viral Immunology and Vaccines, CERVirVac, Zagreb, Croatia.
authors.forename: Beata
authors.initials: B
authors.lastname: Halassy
authors.affiliation: Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, Rockefellerova 10, HR-10 000, Zagreb, Croatia.|Centre of Excellence for Viral Immunology and Vaccines, CERVirVac, Zagreb, Croatia.
authors.forename: Marija
authors.initials: M
authors.lastname: Brgles
mesh_terms: D000818:Animals; D055592:Biophysical Phenomena; D002522:Chlorocebus aethiops; D005374:Filtration; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008458:Measles Vaccine / Q000302:isolation & purification; D008459:Measles virus / Q000737:chemistry / Q000302:isolation & purification*; D009108:Mumps Vaccine / Q000302:isolation & purification; D009109:Mumps virus / Q000737:chemistry / Q000302:isolation & purification*; D014461:Ultracentrifugation; D014709:Vero Cells
chemical_list: D008458:Measles Vaccine; D009108:Mumps Vaccine