The Cell and Gene Therapy (CGT) Catapult established a multi-partner, international consortium to test novel process analytical technologies (PAT) as applied to an exemplar 8-day CAR T-cell culture bioprocess. A critical step in the follow-on analysis of the data generated by the consortium is comparison to robust phenotypic data of the T cell populations.
Multiparameter flow cytometry is a powerful analytical tool that can provide detailed characterisation of complex medicinal cell products and can be used to establish a target product profile (TPP) defining product identity, safety and potency. CGT Catapult partnered with Becton Dickinson (BD) Biosciences, a collaborator on the ‘PAT Consortium’, to develop a complex, 12-colour flow cytometry panel using the BD FACSLyric™ flow cytometer for our 8-day exemplar CAR T-cell bioprocess. In this white paper, we highlight the considerations taken when developing the panel, and importantly, we demonstrated the reliable performance of this panel to identify T cell phenotypes (naïve, memory, activation and exhaustion profiles) over an 8-day culture period. The data generated form part of a larger pool of analytical data that will be used to define future process control parameters and quality control models for CAR T-cell bioprocessing.