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Abstract

The present invention discloses a versatile application of a multimer technology for detecting, manufacturing, and profiling chimeric antigen receptor (CAR)-expressing cells. These antigen-multimers exhibit remarkable specificity, sensitivity, and precision in identifying CAR cells. Moreover, they facilitate over 100-fold magnetic enrichment of rare CAR-T cells, enhancing their detectability. Beyond CAR detection, the multimer technology selectively stimulates CAR-expressing cells, both in soluble and surface-bound formats, during CAR-T cell manufacturing. Unlike existing technologies (such as anti-CD28/CD3 magnetic beads) that non-specifically stimulate and expand all T cells, the multimer approach ensures that only CAR-expressing cells are targeted. Consequently, this yields a CAR-T cell product with significantly higher purity, enhancing treatment efficacy. Furthermore, the multimer technology offers flexibility in CAR detection through nucleotide labeling. This enables high-dimensional CAR-T cell profiling via single-cell multi-omics analyses. Antigen multimers can be seamlessly adapted to other CAR systems by switching the antigen ligand.

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