(W1030-06-39) Comprehensive LC-MS/MS Analysis of VSV-GP Lipidome

Purpose: Investigation of potential critical quality attributes (CQAs) is important to ensure quality and safety of pharmaceutical drugs, including advanced modalities like the Vesicular Stomatitis Virus pseudotyped with LCMV glycoprotein (VSV-GP). Lipids play essential roles in the VSV-GP membrane. Thus, discovery of whether lipids are potential CQAs for VSV-GP become essential.

Methods: Lipids are extracted from VSV-GP infected cells and the virus particles that released into the supernatant. We used liquid chromatography–high-resolution tandem mass spectrometry (LC-MS/MS) to perform a lipidomics analysis to identify and quantify the lipid contents in the infected cells and virus particles.

Results: We identified and quantified some major classes of lipids such as phospholipids and neutral lipids are induced in VSV-GP infected cells compared to uninfected cells at the time of extraction. Moreover, major phospholipids are also present in the VSV-GP particles. Phosphatidylinositols, in particular, are abundant in the virus particle (relative to exosome control samples).

Conclusion: To conclude, VSV-GP infection alters host and virus lipidome and leads to a potential important role of lipids to VSV-GP. Additional studies are planned to examine how the phospholipid content varies across batches and, separately, within batches over the course of the virus life cycle. Further investigation on the role of lipids in PK/PD, immunogenicity and biodistribution is also planned. Overall, this study provides advanced insights into potential CQAs for the VSV-GP platform.