There is considerable excitement surrounding the treatment potential of T cell immunotherapies. Despite the clinical success of chimeric antigen receptor (CAR) T cells, there remain challenges associated with routinely offering these products as treatment alternatives. These challenges include the costly manufacturing process relying on lengthy and complex open workflows with high manual labor requirements that influence product variability. This application note describes the details of a robust CAR T cell manufacturing workflow that can be adapted for cGMP compliance in commercial production of CAR T cells. This semi-automated, closed CAR T process achieves 1 × 1010 expanded T cells with more than 80% enhanced green fluorescent protein (eGFP) transduction efficiency across an 8-day manufacturing process.