Fig. 3. Generation of target candidate-specific CAR constructs and evaluation of in vitro functionality.

a Scheme of the generation of our CAR library. The combination of four backbones differing in spacer length with two scFv orientations and four target candidate specificities resulted in 32 constructs. b Average relative target cell killing, cytokine release, marker upregulation, and overall performance (displayed as circle size) of the selected best-performing CARs from the initial screening assays (y axis). The number of replicates is indicated in parentheses behind the construct name (first value = n of killing assays, second value = n of cytokine release assays, third value = n of marker upregulation measurements). c Representative result for the kinetics of BxPC3 target cell killing by the selected CAR constructs. d Representative results showing cytokine release and activation marker expression patterns upon co-culture with BxPC3 target cells. Shown are mean ± s.e.m. (n = 2). Activation marker expression was measured at end point of cytotoxicity assay, cytokine release patterns were measured after 48 h. Source data are provided as a Source Data file.