High-throughput optimization platform for maximizing virus productivity in mammalian cell cultures

Overview

This research program addresses two persistent bottlenecks in mammalian virus manufacturing: low host cell densities and inconsistent batch-to-batch productivity. By establishing a contaminant-free, nutrient-rich environment and using frequent, data-driven monitoring of cell state, viable cell density, nutrient content, and infection competence, the team can determine optimal cell passaging intervals, target densities, and infection and harvest timing. The approach has been validated in both adherent and suspension cultures using chemically-defined, serum- and protein-free media, and is being extended to additional viruses and host cell lines.

Technical specifications

Key capabilities and findings:

• Adherent culture lentivirus production reached 8.27 x 10^2 transduction units per microliter in chemically-defined, serum- and protein-free medium.
• Suspension cultures consistently achieved 5 to 7.4 x 10^6 viable cells/mL at 94.4 to 99.1 percent viability without medium replacement.
• Increasing the target viable cell density from 5.0 to 6.0 x 10^6 cells/mL raised lentiviral gene copies per cell from 4.4 to 184.4, a 42-fold improvement.
• Statistical analysis of brightfield and fluorescent images, transduction assays, and RT-qPCR inform decisions on passaging, target density, infection timing, and harvest timing.
• Future work targets Morbillivirus in human embryonic kidney cells (or a sponsor-selected mammalian host), with scale-up to high-growth suspension culture featuring high mass transfer and low shear, and without medium replacement.
• High-throughput assays for infected cells and RT-qPCR of select mRNA transcripts and viral genomes support optimization across formats and scales.

Technology readiness level

The platform has been experimentally validated on lentivirus and adenovirus in both adherent and suspension mammalian cell cultures, with quantitative productivity gains demonstrated. Current efforts focus on extending the approach to additional viruses and host cell lines, scaling up to high-density suspension production without medium replacement, and refining high-throughput monitoring for broader operational formats and volume scales.