Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus

Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, Germany

Faculty of Biology and Chemistry, University of Giessen, Heinrich-Buff-Ring 17, 35392 Giessen, Germany

Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Project group Bioresources, Winchesterstr. 3, 35394 Giessen, Germany

Author to whom correspondence should be addressed.

Viruses 2019, 11(8), 725; https://doi.org/10.3390/v11080725

Received: 25 June 2019 / Revised: 31 July 2019 / Accepted: 3 August 2019 / Published: 7 August 2019

(This article belongs to the Special Issue Recent Advances in Morbillivirus Vaccine Development and Oncolytic Virotherapy)

View Full-Text Download PDF

Browse Figures

Citation Export

Abstract

Oncolytic measles virus (MV) is a promising treatment for cancer but titers of up to 10¹¹ infectious particles per dose are needed for therapeutic efficacy, which requires an efficient, robust, and scalable production process. MV is highly sensitive to process conditions, and a substantial fraction of the virus is lost during current purification processes. We therefore conducted forced degradation studies under thermal, pH, chemical, and mechanical stress to determine critical process parameters. We found that MV remained stable following up to five freeze–thaw cycles, but was inactivated during short-term incubation (< 2 h) at temperatures exceeding 35 °C. The infectivity of MV declined at pH < 7, but was not influenced by different buffer systems or the ionic strength/osmolality, except high concentrations of CaCl₂ and MgSO₄. We observed low shear sensitivity (dependent on the flow rate) caused by the use of a peristaltic pump. For tangential flow filtration, the highest recovery of MV was at a shear rate of ~5700 s⁻¹. Our results confirm that the application of forced degradation studies is important to identify critical process parameters for MV purification. This will be helpful during the early stages of process development, ensuring the recovery of high titers of active MV particles after purification. View Full-Text

Keywords: temperature; pH; buffer; ionic strength; osmolality; isoelectric point; shear stress; Measles virus; stability

▼ Show Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited

Share and Cite

MDPI and ACS Style

Loewe, D.; Häussler, J.; Grein, T.A.; Dieken, H.; Weidner, T.; Salzig, D.; Czermak, P. Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus. Viruses 2019, 11, 725. https://doi.org/10.3390/v11080725

AMA Style

Loewe D, Häussler J, Grein TA, Dieken H, Weidner T, Salzig D, Czermak P. Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus. Viruses. 2019; 11(8):725. https://doi.org/10.3390/v11080725

Chicago/Turabian Style

Loewe, Daniel, Julian Häussler, Tanja A. Grein, Hauke Dieken, Tobias Weidner, Denise Salzig, and Peter Czermak. 2019. "Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus" Viruses 11, no. 8: 725. https://doi.org/10.3390/v11080725

Find Other Styles

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Abstract views Pdf views Html views

Article Menu

Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus

Abstract

Share and Cite

Article Metrics

Article Access Statistics

Article Access Map by Country/Region

Further Information

Guidelines

MDPI Initiatives

Follow MDPI