Background & Aim There is a growing need to close and automate chimeric antigen receptor (CAR) T cell therapy manufacturing. CAR T workflows depend on an ex vivogene transfer step for therapeutic efficacy. To streamline this step while keeping flexibility during process development, we introduced a stand-alone protocol software that uses existing Sepax C-Pro technology. SpinOculation C-Pro protocol software automates lentiviral vector (LVV) transduction and maintains a functionally closed system within a single-use disposable kit, and no viral entry enhancers are required. We present different user cases highlighting the flexibility of the SpinOculation C-Pro protocol software parameters to influence process development of LVV transduction efficiencies in combination with application-specific upstream processes and viral source. Methods, Results & Conclusion Methods Case 1 used frozen apheresis units from one healthy donor. Case 2 used fresh apheresis units from three healthy donors. Case 1 included thawing and washing of the apheresis unit with closed, automated processes.In both cases, T cell enrichment and activation were performed using manual, open processes on the first day of the workflow. Activation was continued in T-flask culture for the next 24 hours. Then, on culture day 1 in both cases, cells were split to perform parallel SpinOculation C-Pro process and manual transduction processes in static or shake flask for case 1 and in static flask for case 2. Table 2 shows the SpinOculation C-Pro protocol software parameters used in the two cases. Conclusion As these user cases demonstrate, SpinOculation C-Pro protocol software offers a solution for a closed, automated LVV transduction process step where transduction efficiency can be comparable to a manual, open process. The open, flexible software parameters provide a wide potential for process development and optimization by allowing users to vary multiple parameters . It is important to consider that many factors influence the performance that can be achieved. Notably, transduction efficiency is highly dependent on LVV source and the MOI. Moreover, the upstream preparation of T cells, as well as the method and reagents used to expand transduced T cells, can impact performance.SpinOculation C-Pro protocol software completes our solutions to meet the growing need for fully automated workflow solutions for chimeric antigen receptor T cell therapies to help standardize processes and reduce the impact of donor variability. There is a growing need to close and automate chimeric antigen receptor (CAR) T cell therapy manufacturing. CAR T workflows depend on an ex vivogene transfer step for therapeutic efficacy. To streamline this step while keeping flexibility during process development, we introduced a stand-alone protocol software that uses existing Sepax C-Pro technology. SpinOculation C-Pro protocol software automates lentiviral vector (LVV) transduction and maintains a functionally closed system within a single-use disposable kit, and no viral entry enhancers are required. We present different user cases highlighting the flexibility of the SpinOculation C-Pro protocol software parameters to influence process development of LVV transduction efficiencies in combination with application-specific upstream processes and viral source. Case 1 used frozen apheresis units from one healthy donor. Case 2 used fresh apheresis units from three healthy donors. Case 1 included thawing and washing of the apheresis unit with closed, automated processes.In both cases, T cell enrichment and activation were performed using manual, open processes on the first day of the workflow. Activation was continued in T-flask culture for the next 24 hours. Then, on culture day 1 in both cases, cells were split to perform parallel SpinOculation C-Pro process and manual transduction processes in static or shake flask for case 1 and in static flask for case 2. Table 2 shows the SpinOculation C-Pro protocol software parameters used in the two cases.