With five chimeric antigen receptor (CAR) T cell immunotherapy products currently approved by the FDA for the treatment of hematological cancers, there are increasing efforts to develop novel and better manufacturing technologies and processes for cell therapies to improve efficacy, reduce variability, and reduce cost. To address this possibility, we demonstrate CAR-T manufacturing using the Erbi Breez closed sterile single-use perfusion bioreactor, which operates at a 2 mL working volume and achieve densities in excess of 100e6 cells/mL. We have demonstrated that the perfusion system can perform in-place activation, transduction, and expansion. We show that with improved media exchange rates, cell expansion performance is improved, achieving nearly patient dose levels with more than 400-fold expansion from 0.6 million cells at the point of transduction to more than 200 million cells at 14 days post-transduction at consistently high cell viability of above 95%. These data show that the Breez platform is well suited for cell therapy process development and process characterization studies, and is promising for the production of a cell dosage that is almost sufficient for a patient infusion.