Cell hybrids as virus packaging cells for high efficiency production of gene therapy vectors and viral vaccines

Inventors

Forman, Lawrence • Ngo, Kathy

Assignees

CHO Plus Inc

Abstract

This disclosure provides a technology for adapting host cells to maximize production and improve quality of viral vectors and particles. Cell hybrids are formed from parental cell lines, and divided or cloned into multiple aliquots for testing. Aliquots are chosen that have high production capacity and phenotypic features for virus production, such as an optimal level of intracellular organelles, and used to establish producer cell lines. The producer cells can be genetically altered to express a transgene that encodes viral elements for production of the viral vectors or particles with a therapeutic payload. The hybrid producer cells generate more viral vectors or particles per cell with higher functional titer, thereby lowering the cost of production of pharmaceutical agents for use in gene therapy and immunization.

Core Innovation

The invention provides a process for establishing a packaging cell line for high efficiency production of viral vectors or particles. It uses a starter population of cultured cells that are able to package a species of viral vector or particle when genetically altered to express genes encoding the viral vector or particle. Hybrid cells are formed from the starter population so that each hybrid cell comprises two or more cells, which are then divided into a plurality of aliquots.

Cells from each aliquot are sampled and genetically altered to express genes encoding a test viral vector or particle of the same species. Production of the test viral vector or particle by cells in each aliquot is measured to identify which aliquots contain cells producing a higher functional titer than cells in other aliquots. Cells from one or more identified aliquots are expanded in culture to establish a packaging cell line that, when genetically altered, produces more viral vector or particle per liter of culture fluid than the starter population.

The description further links identifying higher-producing hybrid-cell aliquots to functional intracellular and phenotype-linked features, including intracellular organelles and trafficking-related characteristics. In particular, it emphasizes selection using mitochondria content and reactive oxygen species phenotypes, together with measurement of viral production functional titer and full-to-empty capsids ratio where applicable. It also frames genetic engineering and characterization approaches for producing and selecting high producer cell populations, and it describes that therapeutic payloads may include nucleic acids and proteins, including SARS-CoV-2 antigen examples.

Claims Coverage

The only independent claim is directed to a process for establishing a packaging cell line for high efficiency production of viral vectors or particles. The inventive features center on forming hybrid cells, partitioning into aliquots, genetically altering sampled aliquot cells with a test viral vector or particle, measuring production by functional titer to identify higher-producing aliquots, and expanding selected aliquot cells to establish a packaging cell line with increased production per liter.

Overall, the claim coverage is centered on an aliquot-based hybrid-cell screening workflow that uses genetic alteration with a test viral vector or particle and selection by measured functional titer, followed by expansion of the selected aliquot cells to create a packaging cell line producing more viral vector or particle per liter than the starter population.

Stated Advantages

Documented Applications