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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 CEA (Fl1-39) h(28ζ), which is constructed for the engineering of T cells to target human CEA. The T cells are genetically modified through transduction with a nonviral vector expressing scFv of anti-CEA antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of Colon Cancer.
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CAR Construction :
Fig.2 The CEA binding activity of serially diluted 45scFvLCHα was evaluated by a sandwich ELISA. Antigen binding activity of isolated 45scFvLCHα. ARAKAWA, F., HARUNO, M., KUROKI, M., KANDA, H., WATANABE, T., MISUMI, Y., & MATSUOKA, Y. (1993). Construction and Expression of Two Mouse—Human Chimeric Antibodies with High Specificity and Affinity for Carcinoembryonic Antigen. Hybridoma, 12(4), 365-379. |
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CAR Construction :
Fig.3 Competitive inhibition assays of chimeric antibodies against parental MAbs for CEA binding in SPEIAs using purified biotinylated CEA. Fl 1-39 was dried onto wells of 96-well plates. ARAKAWA, F., HARUNO, M., KUROKI, M., KANDA, H., WATANABE, T., MISUMI, Y., & MATSUOKA, Y. (1993). Construction and Expression of Two Mouse—Human Chimeric Antibodies with High Specificity and Affinity for Carcinoembryonic Antigen. Hybridoma, 12(4), 365-379. |
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CAR Construction :
Fig.4 Reactivity of mouse MAbs (Fll-35 and Fll-39) with CEA and CEA-related antigens. A SPEIAs using antigen-coated plates and a biotinylated goat anti-human y-chain antibody or a biotinylated horse anti-mouse IgG(H+L) antibody. ARAKAWA, F., HARUNO, M., KUROKI, M., KANDA, H., WATANABE, T., MISUMI, Y., & MATSUOKA, Y. (1993). Construction and Expression of Two Mouse—Human Chimeric Antibodies with High Specificity and Affinity for Carcinoembryonic Antigen. Hybridoma, 12(4), 365-379. |
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CAR Construction :
Fig.1 Detection of the expressed F39scFv/CIR receptors in the transfectoma by flow cytometry. Cell surface expression of the F39scFv CIR receptor protein in the transfectoma was demonstrated by FACS, Arakawa, F., Shibaguchi, H., Xu, Z. H. O. N. G. W. E. I., & Kuroki, M. (2002). Targeting of T cells to CEA-expressing tumor cells by chimeric immune receptors with a highly specific single-chain anti-CEA activity. Anticancer research, 22(6C), 4285-4289. |
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