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.

Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium

“…MV particles are large (~350 nm, range 50-1000 nm [16]) and complex structures, and their inherent instability demands well-chosen DSP strategies [9,17,18]. However, few MV studies have focused on DSP.…”

Screening different host cell lines for the dynamic production of measles virus

“…MV production is sensitive to several process parameters including temperature and pH, which means that poorly‐controlled processes are unstable and non‐reproducible . The production of MV in a STR provides the opportunity for total process monitoring and control, but even when critical process parameters were maintained within strict tolerances (± 0.1 pH and ± 0.5°C) we found that the production of MV using Vero cells resulted in variable virus production kinetics.…”

High titer oncolytic measles virus production process by integration of dielectric spectroscopy as online monitoring system

“…However, neither process can yet provide the concentrations of active Measles virus required for therapeutic applications. Although enveloped viruses in general are sensitive to changes in osmolality, some reports indicate that Measles virus is comparatively robust, whereas others confirm its sensitivity (Sviben et al, 2016;Weiss et al, 2013). Measles viruses are sensitive to low pH and UV light, as is typical for enveloped viruses (Grein, Michalsky, & Czermak, 2014;Sviben, Forčić, Kurtović, Halassy, & Brgles, 2016;Weiss et al, 2013).…”