| Products/Services Used | Details | Operation |
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| Protein and Antibody Isolation> | An FCM analysis was used to detect anti-HER2 scFv on CAR-T cells and aPD-L1 scFv on POMVs. CAR-T cells were washed with PBS containing 1% BSA, incubated at 4 °C with biotinylated protein L at 1 mg ml−1 (CAS no. M00097, GenScript), washed, and stained with phycoerythrin-conjugated streptavidin (CAS no. 405203, BioLegend). For POMVs, vesicles were bound to 4-μm aldehyde/sulfate latex beads (CAS no. A37304, ThermoFisher), washed and stained at 4 °C with a phycoerythrin-conjugated anti-Myc antibody or isotype control (CAS no. ab18443, Abcam) to detect Myc-tagged aPD-L1 scFv. | Get A Quote |
Major challenges facing chimeric antigen receptor (CAR)-T cell therapy for solid tumours include the immunosuppressive tumour microenvironment and the heterogeneity of antigen expression. Bacterial outer membrane vesicles (OMVs) naturally activate the immune system and can be engineered for drug delivery. Here we develop a bacterial OMV-based immunosuppression reversal and optimized antigen decoration platform for CAR-T therapy (BROAD-CAR), in which OMVs are modified to express a high-affinity anti-PD-L1 antibody and load plasmids encoding the target antigen for CARs. By blocking the PD-1/PD-L1 signalling pathway, our tumour-targeting platform enhances the antitumour activity of CAR-T cells both in vitro and in... More