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Recombinant Moloney murine leukemia virus (MMLV) retroviral vectors serve as efficient viral vector tools for introducing genes permanently into a wide variety of dividing cells, and provide an alternative to modify primary T cells. Creative biolabs has developed retroviral CAR vector pMMLV ROR1 (ERR1-TOP43) h(28ζ), which is constructed for the engineering of T cells to target human ROR1. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-ROR1 antibody linked to a CD28 transmembrane domain and CD3-zeta signaling domains. And the vector product was designed for the treatment of Cancer.
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CAR Construction :
Fig.1 Cell surface binding of bispecific ROR1 × CD3 scFv-Fc. The indicated bispecific ROR1 × CD3 biAbs were analyzed for binding to human CD3-positive T cell line Jurkat and human ROR1-positive cell lines K562/ROR1 and JeKo-1 by flow cytometry at a concentration of 5 μg/mL followed by APC-conjugated goat Qi, J., Li, X., Peng, H., Cook, E. M., Dadashian, E. L., Wiestner, A., ... & Rader, C. (2018). Potent and selective antitumor activity of a T cell-engaging bispecific antibody targeting a membrane-proximal epitope of ROR1. Proceedings of the National Academy of Sciences, 115(24), E5467-E5476. |
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CAR Construction :
Fig.2 Cross-linking of 5 × 104 primary human T cells (stained with CellTrace Far Red) and 5 × 104 K562/ROR1 cells (stained with CellTrace CFSE) in the presence of 1 μg/mL ROR1 × CD3 and control scFv-Fc. Double-stained events were detected by flow cytometry. Qi, J., Li, X., Peng, H., Cook, E. M., Dadashian, E. L., Wiestner, A., ... & Rader, C. (2018). Potent and selective antitumor activity of a T cell-engaging bispecific antibody targeting a membrane-proximal epitope of ROR1. Proceedings of the National Academy of Sciences, 115(24), E5467-E5476. |
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CAR Construction :
Fig.3 The cytotoxicity of ROR1 × CD3 scFv-Fc was tested with ex vivo expanded primary human T cells and K562 cells. Shown are mean ± SD values from three independent triplicates. Qi, J., Li, X., Peng, H., Cook, E. M., Dadashian, E. L., Wiestner, A., ... & Rader, C. (2018). Potent and selective antitumor activity of a T cell-engaging bispecific antibody targeting a membrane-proximal epitope of ROR1. Proceedings of the National Academy of Sciences, 115(24), E5467-E5476. |
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