Figure 6.
Tandem CAR design has impaired antigen binding compared with the monospecific CARs. (A) CAR expression determined by flow cytometry staining with anti-STII, CD19-biotin, and CD79ab-Fc staining on sorted EGFRt+ T cells. Right panels show quantification of normalized gMFI determined from flow cytometric staining of 5 different donors. Quantification was normalized to the mean obtained by staining monospecific CD19 CAR T cells with anti-STII or CD19-biotin binding. Quantification for CD79ab-Fc binding was normalized to that obtained with the monospecific CD79a CAR. (B) Flow cytometric histograms of indicated CAR T cells gated on EGFRt+ cells, showing anti-MYC or CD79ab-Fc staining. Right panel shows quantification of normalized gMFI flow cytometric staining of 3 different donors. Quantification was normalized to CD79a CAR T cell for anti-MYC or CD79ab-Fc binding for each donor. Indicated P values were with ∗∗∗P < .001, ∗∗P < .01, ∗P <.05 or ns were calculated with one-way ANOVA Tukey multiple comparisons test and panel B paired t test. ns, not significant.

Tandem CAR design has impaired antigen binding compared with the monospecific CARs. (A) CAR expression determined by flow cytometry staining with anti-STII, CD19-biotin, and CD79ab-Fc staining on sorted EGFRt+ T cells. Right panels show quantification of normalized gMFI determined from flow cytometric staining of 5 different donors. Quantification was normalized to the mean obtained by staining monospecific CD19 CAR T cells with anti-STII or CD19-biotin binding. Quantification for CD79ab-Fc binding was normalized to that obtained with the monospecific CD79a CAR. (B) Flow cytometric histograms of indicated CAR T cells gated on EGFRt+ cells, showing anti-MYC or CD79ab-Fc staining. Right panel shows quantification of normalized gMFI flow cytometric staining of 3 different donors. Quantification was normalized to CD79a CAR T cell for anti-MYC or CD79ab-Fc binding for each donor. Indicated P values were with ∗∗∗P < .001, ∗∗P < .01, ∗P <.05 or ns were calculated with one-way ANOVA Tukey multiple comparisons test and panel B paired t test. ns, not significant.

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