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. 2020 May 8;11:2283. doi: 10.1038/s41467-020-16160-5

Fig. 1. High-throughput generation and screening of BCMA/CS1 OR-gate CARs.

Fig. 1

a A vertically integrated optimization process for CAR-T cell therapy development. b Schematic of a single-chain bispecific (OR-gate) CAR, which contains two ligand-binding domains connected in tandem. A panel of CAR variants was constructed from two CS1-binding scFvs and three BCMA-specific binding domains. Both murine and humanized versions of BCMA-binding scFvs (c11D5.3 and J22.9-xi) were evaluated. The CS1-binding huLuc63 and Luc90 scFvs were fixed at the membrane-distal and membrane-proximal positions, respectively, based on binding-epitope analysis. c Methodology for producing and screening bispecific CARs. CAR-T cells were generated in a 14-day cycle. Starting on day 9 post-stimulation, CAR-T cells were characterized for antitumor function in various assays, which could last up to 2 weeks.