Absolute quantification of viral proteins from pseudotyped VSV-GP using UPLC-MRM

The development of oncolytic viral therapy has gained considerable momentum in recent years. Vesicular stomatitis virus glycoprotein (VSV-GP) is a new biotherapeutic emerging in the oncolytic viral therapy platform. Novel analytical assays that can accurately and precisely quantify the viral protein...

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Published inMicrobiology spectrum Vol. 12; no. 8; p. e0365123
Main Authors Basu, Rajeswari, Dambra, Richard, Jiang, Di, Schätzlein, Sophia A., Njiyang, Shu, Ashour, Joseph, Chiramel, Abhilash I., Vigil, Adam, Papov, Vladimir V.
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 06.08.2024
Subjects
absolute quantification
Genomics and Proteomics
GPC tracking
oncolytic virus
Research Article
GPC tracking
oncolytic virus
absolute quantification
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Summary:The development of oncolytic viral therapy has gained considerable momentum in recent years. Vesicular stomatitis virus glycoprotein (VSV-GP) is a new biotherapeutic emerging in the oncolytic viral therapy platform. Novel analytical assays that can accurately and precisely quantify the viral proteins are a necessity for the successful development of viral vector as a biotherapeutic. We developed an ultra-high performance liquid chromatography multiple reaction monitoring-based assay to quantify the absolute concentrations of the different structural proteins of VSV-GP. The complete processing of GP complex (GPC) is a prerequisite for the infectivity of the virus. The assay extends the potential for quantifying full-length GPC, which provides an understanding of the processing of GPC (along with the quantification of GP1 and GP2 separately). We used this assay in tracking GPC processing in HEK-293-F production cell lines infected with VSV-GP.
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Present address: Denali Therapeutics Inc., Development Sciences, South San Francisco, California, USA
All authors were employed by Boehringer Ingelheim Pharmaceuticals Inc. (BIPI) at the time of the experiments.
Present address: Regeneron Pharmaceuticals, Tarrytown, New York, USA
The authors declare no conflict of interest.
ISSN:2165-0497
2165-0497
DOI:10.1128/spectrum.03651-23