Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus
Oncolytic measles virus (MV) is a promising treatment for cancer but titers of up to 10<sup>11</sup> 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...
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doaj-bf7221812fc2415aac6906d67ee334ae2020-11-25T00:28:02ZengMDPI AGViruses1999-49152019-08-0111872510.3390/v11080725v11080725Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles VirusDaniel Loewe0Julian Häussler1Tanja A. Grein2Hauke Dieken3Tobias Weidner4Denise Salzig5Peter Czermak6Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, GermanyOncolytic measles virus (MV) is a promising treatment for cancer but titers of up to 10<sup>11</sup> 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 CaCl<sub>2</sub> and MgSO<sub>4</sub>. 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<sup>−1</sup>. 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.https://www.mdpi.com/1999-4915/11/8/725temperaturepHbufferionic strengthosmolalityisoelectric pointshear stressMeasles virusstability |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Daniel Loewe Julian Häussler Tanja A. Grein Hauke Dieken Tobias Weidner Denise Salzig Peter Czermak |
spellingShingle |
Daniel Loewe Julian Häussler Tanja A. Grein Hauke Dieken Tobias Weidner Denise Salzig Peter Czermak Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus Viruses temperature pH buffer ionic strength osmolality isoelectric point shear stress Measles virus stability |
author_facet |
Daniel Loewe Julian Häussler Tanja A. Grein Hauke Dieken Tobias Weidner Denise Salzig Peter Czermak |
author_sort |
Daniel Loewe |
title |
Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus |
title_short |
Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus |
title_full |
Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus |
title_fullStr |
Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus |
title_full_unstemmed |
Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus |
title_sort |
forced degradation studies to identify critical process parameters for the purification of infectious measles virus |
publisher |
MDPI AG |
series |
Viruses |
issn |
1999-4915 |
publishDate |
2019-08-01 |
description |
Oncolytic measles virus (MV) is a promising treatment for cancer but titers of up to 10<sup>11</sup> 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 CaCl<sub>2</sub> and MgSO<sub>4</sub>. 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<sup>−1</sup>. 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. |
topic |
temperature pH buffer ionic strength osmolality isoelectric point shear stress Measles virus stability |
url |
https://www.mdpi.com/1999-4915/11/8/725 |
work_keys_str_mv |
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