Journal: Biomedicines

Article Title: Evaluation of Anti-CAR Linker mAbs for CAR T Monitoring after BiTEs/bsAbs and CAR T-Cell Pretreatment

doi: 10.3390/biomedicines12081641

Figure Lengend Snippet: Anti-CAR linker mAbs allows for specific and sensitive CAR T monitoring in patients treated with any clinically approved CAR T therapies except Cilta-cel. ( A ) The structures of BCMA- and CD19-specific CARs are depicted in the illustration. CD19 antigen-specific CARs: Brexu-cel, Liso-cel, Axi-cel, and Tisa-cel; BCMA-specific CARs: Ide-cel and Cilta-cel. ( B ) Flow cytometry-based CAR detection analyses are shown, performed with biotinylated anti-CAR linker mAbs and APC-conjugated anti-biotin Abs to all approved BCMA and CD19 CAR T-cell products using isolated lymphocyte specimens from patients’ blood (n = 3). ( C ) Flow cytometry-based CAR detection analyses are shown, performed with Alexa 647-conjugated anti-CAR linker mAbs to all approved BCMA and CD19 CAR T-cell products using isolated lymphocyte specimens from patients’ blood (n = 3). The CAR-positive cells were determined from the CD3-positive cells and significances calculated based on BCMA- and CD19-antigen-based CAR detection reagent (Miltenyi = 100%). Statistical analysis was performed using Student’s t -test. * p ≤ 0.05%; ** p ≤ 0.01%; *** p ≤ 0.001%.

Article Snippet: Anti-G 4 S CAR linker rabbit mAb (E7O2V), anti-Whitlow CAR linker rabbit mAb (E3U7Q) (biotinylated or Alexa Fluor ® 647-conjugated), and rabbit IgG XP ® isotype-Alexa Fluor ® 647 (DA1E) were obtained from Cell Signaling Technology Europe B.V. (Leiden, The Netherlands).

Techniques: Flow Cytometry, Isolation