Figure 2
Figure 2. CD22 CARs expressing high affinity scFv (HA22), standard affinity (BL22), and membrane proximal binding (m971) anti-CD22 scFv-derived domains. (A) Second-generation CAR constructs (CD28 and CD3zeta or 4-1BB and CD3zeta signaling domains) and third-generation constructs (CD28, 4-1BB, and CD3zeta) expressed HA22, BL22, m971, or FMC63-derived scFv, ± an IgG1-derived CH2CH3 spacer domain. (B) HA22 and BL22 scFvs bind Ig domain 3, whereas m971 binds within Ig domains 5-7 of CD22. (C) Expression on transduced T cells of CD22 CARs containing CH2CH3 domains (top row) or in the shorter format, omitting this domain (bottom row). CD22 CARs were detected by CD3-APC (y-axis) and CD22-Fc followed by an anti–IgG-Fc-FITC stain. Percent transduction is noted in the top-right quadrant of each plot. (D) MFI, y-axis, of the indicated CD22 CAR constructs, x-axis. Results are representative of more than 3 retroviral supernatant preparations.

CD22 CARs expressing high affinity scFv (HA22), standard affinity (BL22), and membrane proximal binding (m971) anti-CD22 scFv-derived domains. (A) Second-generation CAR constructs (CD28 and CD3zeta or 4-1BB and CD3zeta signaling domains) and third-generation constructs (CD28, 4-1BB, and CD3zeta) expressed HA22, BL22, m971, or FMC63-derived scFv, ± an IgG1-derived CH2CH3 spacer domain. (B) HA22 and BL22 scFvs bind Ig domain 3, whereas m971 binds within Ig domains 5-7 of CD22. (C) Expression on transduced T cells of CD22 CARs containing CH2CH3 domains (top row) or in the shorter format, omitting this domain (bottom row). CD22 CARs were detected by CD3-APC (y-axis) and CD22-Fc followed by an anti–IgG-Fc-FITC stain. Percent transduction is noted in the top-right quadrant of each plot. (D) MFI, y-axis, of the indicated CD22 CAR constructs, x-axis. Results are representative of more than 3 retroviral supernatant preparations.

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