All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.
Sleeping Beauty (SB) transposon, a type of nonviral integrative vectors, provides an alternative to modify primary T cells. Creative biolabs has developed SB transposon CAR vector pSBCAR1 CD22 (HA22) h(28OXζ), which is constructed for the engineering of T cells to target human CD22. The T cells are genetically modified through transduction with a nonviral vector expressing scFv of anti-CD22 antibody linked to a CD28 transmembrane domain/ endodomain and OX40, CD3-zeta signaling domains. And the vector product was designed for the treatment of Acute lymphoblastic leukemia (ALL).
CAR Construction : HA22-CD28-CD3ζ, HA22-CD28-41BB-CD3ζ Fig.1 Expression on transduced T cells of CD22 CARs in the second and third-generation constructs. 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. Haso, W., Lee, D. W., Shah, N. N., Stetler-Stevenson, M., Yuan, C. M., Pastan, I. H., ... & Orentas, R. J. (2013). Anti-CD22–chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood, The Journal of the American Society of Hematology, 121(7), 1165-1174. |
CAR Construction : HA22-CD28-41BB-CD3ζ Fig.2 HA22 and BL22-derived CD22 CARs mediate lytic activity. The lytic activity HA22 and BL22-derived CD22 CARs was compared against ALL cell lines at the indicated E:T ratios in a 4-hour 51Cr-release assay. Haso, W., Lee, D. W., Shah, N. N., Stetler-Stevenson, M., Yuan, C. M., Pastan, I. H., ... & Orentas, R. J. (2013). Anti-CD22–chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood, The Journal of the American Society of Hematology, 121(7), 1165-1174. |
CAR Construction : Fig.3 Interaction between CD22 and HEK 293T cell surface-displayed scFv Four individual clones (PT.1, PT.2, PT.3, and PT.4) of scFvs with the same mutation (PT) isolated from a single-pass enrichment were tested for the scFv expression (scFv) on HEK 293T cells with anti-myc mAb detected by FITC and for the binding to the antigen with CD22-Fc (CD22) detected by PE. Ho, M., Nagata, S., & Pastan, I. (2006). Isolation of anti-CD22 Fv with high affinity by Fv display on human cells. Proceedings of the National Academy of Sciences, 103(25), 9637-9642. |
CAR Construction : Fig.4 Equilibrium binding titration curves to determine dissociation constants, KD. MFI (%) of PE is plotted versus the various concentrations of biotinylated CD22-Fc used to label surface-displayed BL22 (WT, KD = 5.8 nM, Bmax = 453), HA22 (KD = 2.5 nM, Bmax = 293), or mutant PT (KD = 1.2 nM, Bmax = 275) antibody. Ho, M., Nagata, S., & Pastan, I. (2006). Isolation of anti-CD22 Fv with high affinity by Fv display on human cells. Proceedings of the National Academy of Sciences, 103(25), 9637-9642. |
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