Prior work has demonstrated scalability of the Gibco CTS AAV‑MAX Helper‑Free AAV Production System to produce AAV in the Thermo Scientific DynaDrive Single‑Use Bioreactor (S.U.B.) at the 50 L scale [1]. As further continuation of that work, this study showcases scaling this same process up to a commercial manufacturing scale of 1,000 L working volume in the 5,000 L DynaDrive S.U.B., along with specific guidance on how to successfully perform these processes at this scale.