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Performance comparison of domestic and imported CD3/CD28 activation beads for CAR-T cell manufacturing
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Abstract:
[Abstract] Objective: To compare the performance differences of domestic and imported CD3/CD28 activation beads for manufacturing CAR-T cells, providing a backup or alternative for domestic CAR-T cell research and manufacture. Methods: A mature protocol using imported CD3/CD28 activation beads with a 1∶1 ratio for CD3+ T cells was implemented as research control. Domestic beads were used with gradient ratios of 1∶2, 1∶1, and 2∶1 to activate T cells. 72 h after T cell activation, CAR-T cells were manufactured by CAR lentiviral infection and cell proliferation was monitored at 2-, 4-, and 7-days post-infection. Flow Cytometry was used to detect CAR-T cell positivity 5 days after infection and to detectCD4/CD8 phenotype of CAR-T cells and PD1+ TIM3+ cell exhaustion ratio 8 days after infection. Results: CAR-T cells manufactured by domestic CD3/CD28 activation beads exhibited similar phenotype compared with those manufactured by imported CD4/CD8 beads. The positive rate of CAR-T cells prepared with domestic beads was slightly lower than that of imported beads (53.7% vs 57.9%). However, the proliferation of CAR-T cells manufactured by domestic beads was about twice that of cells manufactured by imported beads, and the exhaustion level was only half that of imported beads (4.21% vs 7.91%). Moreover, the use of domestic magnetic beads was lower than that of imported magnetic beads, which was advantageous for cutting the costs of CAR-T cells research and manufacture. Conclusion: Domestic CD3/CD28 activation beads used for CAR-T cells manufacturing demonstrate comparable overall performance to their imported counterparts, showing potential as a backup or alternative for imported beads.
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