Risk analysis of leachables in cell and gene therapy using a CAR-T model process.

With the rapidly emerging field of autologous therapies, Single-Use (SU) technologies are increasingly used in personalized medicine due to their manifold advantages. Although qualification of the starting material of autologous therapies such as the CAR-T process has been highlighted, little attention has been paid to the effect of leachables on cell-based therapies, even if recent studies indicate interactions of leachables with cells. To close this gap, this study presents a risk-analysis of SU-material on a CAR-T process and identifies hazards imposed by tubing materials and leachables thereof. In order to represent a CAR-T process in its entirety, two test systems, namely a lentivirus production process and primary T-cells, were used. While the effects on lentivirus production are comparable to those reported for antibody production processes in CHO cells, we found that PVC material and corresponding leachables, i.e. plasticizer, inhibit cell growth of primary T-cells to a great extent. Additionally, our results indicate that critical quality attributes are affected by the PVC material.