Modular Cell Line for Scalable and Rapid In Vitro Evaluation of Chimeric Antigen Receptors

Abstract

AbstractThe development of novel Chimeric Antigen Receptor T-cell (CAR-T-cell) products relies on continuousin vitrotesting of new antigen/antibody pairs, as well as modification of CAR composition itself. Since any Tumor-Associated Antigens (TAA) have been chosen and proven to be specific, and before the firstin vivostudies these CAR compositions have to be extensively validatedin vitro. Conventionalin vitrotests rely on TAA expressing immortalized cell lines or immortalized cell lines derivatives engineered to stably express antigen of interest, making this type of model not fully scalable when it comes to multiple TAA/antibody pairs. Therefore, there is a need for a representative, scalable model for preclinical assessment. Here, we introduce a novel model cell line approach that can be easily adapted for different experiments by anchoring ALFA-tagged proteins on the cell surface using an anti-ALFA-tag membrane single-domain antibody. This modular multi-component system has the potential to reduce novel CARin vitroevaluation procedures from months to weeks.