ArticleMicrobiology spectrum2026
Integrating replication kinetics and ultrastructural analysis to identify targets for optimizing rVSV bioproduction.
Article in Microbiology spectrum, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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8 authors.
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Abstract
The growing demand of viral products for gene therapy and as oncolytic agents underscores the importance of optimizing virus production processes in the biopharmaceutical industry. Beyond established bioprocess parameters such as infectious titer, additional analytical methods are required to gain deeper insights into these processes. Electron microscopy, a well-established technique in virology, is rarely utilized in bioprocess design. In this study, we aim to identify potential targets for bioprocess optimization by combining ultrastructural changes of infected producer cells with the replication kinetics of a therapeutic virus. As a model system, we used an oncolytic recombinant vesicular stomatitis virus (rVSV). Replication kinetics were assessed by tissue culture infectious dose (TCID50), quantitative polymerase chain reaction, and optical cytometry. Transmission electron microscopy of high-pressure frozen, freeze-substituted samples was used to analyze the ultrastructure of rVSV-infected cells at different timepoints. Using this approach, we identified three key areas for bioprocess optimization. First, a decrease of the diameter of producer cells over the course of rVSV infection offers potential for online monitoring. Second, clusters of virions attached to the cell membrane of producer cells suggest the presence of cellular restriction factors, which could be overcome by strategies enhancing virus release. Finally, the observation of inclusion bodies surrounded by endoplasmic reticulum indicates a surplus of genomic copies within the producer cells, representing a bottleneck that could be targeted to increase particle assembly after genome replication.
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