Single-step rapid chromatographic purification and characterization of clinical stage oncolytic VSV-GP

Abstract

Purification of viruses, especially for therapeutic purposes, is a tedious and challenging task. The challenges arise due to the size and surface complexity of the virus particles. VSV-GP is a promising oncolytic virus, which has been approved for phase I clinical trials by the Food and Drug Administration (FDA) of United States and Paul Ehrlich Institute (PEI) of Germany. The virus particles of VSV-GP are larger in size than vectors commonly used for gene therapy (e.g., adenovirus, adeno-associated virus, etc.). The current established proprietary clinical-grade manufacturing process for the purification of VSV-GP encompasses several chromatographic and non-chromatographic steps. In this study, we describe a new single-step purification process for the purification of VSV-GP virus, using cation exchange convective flow column with relatively higher yields. The purified virus was characterized for its quality attributes using TCID50 assay (for viral infectivity), host cell protein contaminant ELISA, SDS-PAGE, size exclusion chromatography (SEC), and cryo-electron microscopy. Furthermore, the purified viral therapeutic material was tested in vivo for its efficacy and safety. All these characterization methods demonstrated a therapeutic virus preparation of high purity and yield, which can be readily used for various studies.

Absolute Quantification of Viral Proteins from Pseudotyped Vesicular Stomatitis Virus (VSV-GP) using Ultra High-Performance Liquid Chromatography- Multiple Reaction Monitoring (UPLC-MRM)

“…VSV-GP virus batches were produced in HEK-293F cells in either shaker flask or bioreactor formats (depending on batch ID). After infecting cells at an MOI of 0.0005, virus was purified from clarified harvest via centrifugation and/or chromatographic methods similar to previous reports (58, 59). The downstream processes employed were slightly unique for each batch (for example the use of different purification columns or formulations).…”

A new strategy for treating colorectal cancer: Regulating the influence of intestinal flora and oncolytic virus on interferon

“… 181 Similarly, research tested a IFN-II-engineered VSV variant and found that, compared with VSV, the virus can inhibit cancer and improve immune reaction by decreasing tumor infiltration of Tregs and enhancing tumor infiltration of CTLS. 182 It is worth noting that oncolytic VSV expressing IFN-II is presently in clinical tests on solid tumor, 183 which has broad development prospects ( Table 3 ).…”