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

Measles 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 and basic research of these viruses, the preparation of highly purified and infective viruses is a prerequisite, and to meet that aim, knowledge of their stability and biophysical properties is crucial. Our goal was detailed investigation of stability of MeV and MuV under various pH, different temperatures, shear stress, filtration and storage conditions, as well as the impact of two commonly used purification techniques, ultracentrifugation and diafiltration, which were compared with regard to their efficacy and impact on virus properties. Virus titers were estimated by CCID50 assay, particle size and concentration by Nanoparticle Tracking Analysis (NTA) measurements, and host cell protein content by ELISA. Results demonstrated stability of MuV and MeV under pH 9 and pH above 4 and 5, respectively, while pH > 9 induced aggregation. Storage without stabilizer did not induce structural changes, but the reduction of infectivity during 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 lower impact on virus viability. Diafiltration resulted in higher both total and infective virus particle recovery than ultracentrifugation. These results provide important data for research of all upstream and downstream processes on these two viruses regarding biotechnological production and basic research.

measles virus ; mumps virus ; ultracentrifugation ; diafiltration ; Nanoparticle tracking analysis ; virus stability

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