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

Abstract

Measles virus (MV) has a natural affinity for cancer cells and oncolytic MV preparations have therefore been investigated in several clinical trials as a potential treatment for cancer. The main bottleneck in the administration of oncolytic MV to cancer patients is the production process, because very large doses of virus particles are required for each treatment. Here, we investigated the productivity of different host cells and found that a high infection efficiency did not necessarily result in high virus yields because virus release is also dependent on the host cell. As well as producing large numbers of active MV particles, host cells must perform well in dynamic cultivation systems. In screening experiments, the highest productivity was achieved by Vero and BJAB cells, but only the Vero cells maintained their high virus productivity when transferred to a stirred tank reactor. We used dielectric spectroscopy as an online monitoring system to control the infection and harvest times, which are known to be critical process parameters. The precise control of these parameters allowed us to achieve higher virus titers with Vero cells in a stirred tank reactor than in a static cultivation system based on T-flasks, with maximum titers of up to 10 TCID ml . © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:989-997, 2017.

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

“…Michael Muehlebach (Paul-Ehrlich-Institute, Langen, Germany). The dynamic production of MV in suspension was carried out in a stirred-tank reactor [6,8]. Vero cells (CCL-81, ATCC) were grown on 3 g L −1 Cytodex 1 microcarriers (GE Healthcare, Chicago, Illinois, USA) in SCM or SFM.…”

“…Measles virus (MV) is a leading candidate owing to the excellent safety profile of the measles vaccine [2]. However, at least 10 8 infectious MV particles are needed per dose for cancer therapy [3,4] and this has prompted extensive studies focusing on the optimization of upstream processes [5][6][7][8][9][10]. The deeper understanding of MV production gained from these studies has shown that shear stress and aeration play a key role in process productivity [7].…”

Vero cell upstream bioprocess development for the production of viral vectors and vaccines

“…However, the long-standing experience with the Vero cell line and its acceptance by regulatory authorities, continue to make it the substrate of choice for many vaccine manufacturers ( Barrett et al, 2009 ; Genzel, 2015 ). Moreover, in direct comparison with other cell lines, Vero has shown superior virus productivity for a range of viruses, including dengue virus, enterovirus type 71, Japanese encephalitis virus, measles virus and Peste des Petits ruminants virus ( Grein et al, 2017 ; Liu et al, 2008 ; Silva et al, 2008 ; Wu et al, 2004 ; Wu and Huang, 2000 ). For influenza virus, however, MDCK cells demonstrated superior virus production than Vero cells ( Genzel et al, 2010 ).…”

“…More recently, Vero cells are also investigated as cell substrate for the production of certain oncolytic viruses ( Grein et al, 2017 ). In contrast to infectious disease preventing vaccines, viral oncolytic therapy fights cancer by selectively targeting and killing cancer cells through oncolytic virus replication.…”

“…The demand can be compared to that from gene therapy applications, and is generally more than 1000 times higher than for vaccination ( Weiss et al, 2012 ). For MeV based systems, for example, even 10 7 –10 8 times more MeV particles are required for oncolytic therapy treatment of one patient than for one vaccine dose ( Grein et al, 2017 ). Therefore, reaching higher production yields is even more crucial for the implementation and establishment of this new treatment technology.…”