Cell Isolation:Article Title: Tumor antigens preferentially derive from unmutated genomic sequences in melanoma and non-small cell lung cancer
Article Snippet: CD8 + T cells from D50 (leukapheresis product from a healthy HLA-A*02:01-positive donor from BioIVT) were expanded in eight different conditions: with individual aeTSAs (VLMMKLEDL, RLLELHITM, VLWRGDSPL, YQIGQVQGV), control peptides (MelanA26-35A27L, ELAGIGILTV; pp65495–503, NLVPMVATV), an aeTSA from ovarian cancer (Ov-aeTSA, LLSSKLLLM) described as immunogenic by our group , all presented by HLA-A*02:01, or DMSO as the negative control. .. Thawed PBMCs were enriched for naive CD8 + T cells using the Human CD8 + T Cell Isolation Kit combined with CD45RO and CD57 MicroBeads (Miltenyi Biotec) to remove memory T cells from the CD8 + T cell fraction. .. These naive CD8 + T cells were then expanded for 14 d with T Cell TransAct (Miltenyi Biotec) in TexMACS Medium (Miltenyi Biotec) supplemented with interleukin (IL)-7 and IL-21 (PeproTech, at final concentrations of 5 ng ml −1 and 10 ng ml −1 , respectively) before specific expansion.
Article Title: CDK4/6 inhibition reprograms the breast cancer immunopeptidome via Rb-dependent chromatin and transcriptomic remodeling.
Article Snippet: 24 Cell Reports 44, 116676, December 23, 2025 Gene expression and GSEA analyses Transcriptomic data generated in this study, as well as data obtained from GSE157383 58 (MCF7, MDAMB453, MDAMB468 data) and GSE241943 (MCF7 xenografts data), were all aligned with STAR (version 2.7.1) 123 on the GRCh38 (gene annotation from Gencode version 33, based on Ensembl 99). .. Thawed PBMCs were enriched for naive CD8 + T cells using the Human CD8 + T cell Isolation Kit combined with CD45RO and CD57 MicroBeads (Miltenyi Biotec) to deplete memory T cells. .. These naive CD8 + T cells were then expanded for 14 days with T cell TransAct (Miltenyi Biotec) in TexMACS Medium (Miltenyi Biotec) supplemented with interleukin-7 (5 ng/mL, Thermo Fisher Scientific) and interleukin-21 (10 ng/mL, PeproTech).
Article Title: Tumor antigens preferentially derive from unmutated genomic sequences in melanoma and non-small cell lung cancer.
Article Snippet: CD8+ T cells from D50 (leukapheresis product from a healthy HLA-A*02:01-positive donor from BioIVT) were expanded in eight different conditions: with individual aeTSAs (VLMMKLEDL, RLLELHITM, VLWRGDSPL, YQIGQVQGV), control peptides (MelanA26-35A27L, ELAGIGILTV; pp65495–503, NLVPMVATV), an aeTSA from ovarian cancer (Ov-aeTSA, LLSSKLLLM) described as immunogenic by our group92, all presented by HLA-A*02:01, or DMSO as the negative control. .. Thawed PBMCs were enriched for naive CD8+ T cells using the Human CD8+ T Cell Isolation Kit combined with CD45RO and CD57 MicroBeads (Miltenyi Biotec) to remove memory T cells from the CD8+ T cell fraction. .. These naive CD8+ T cells were then expanded for 14 d with T Cell TransAct (Miltenyi Biotec) in TexMACS Medium (Miltenyi Biotec) supplemented with interleukin (IL)-7 and IL-21 (PeproTech, at final concentrations of 5 ng ml−1 and 10 ng ml−1, respectively) before specific expansion.
Imaging:Article Title: Inhibition of autophagy enhances the antitumor efficacy of T/CAR T cell against neuroblastoma
Article Snippet: Fluorochromes were photobleached or released using REAlease technology (Miltenyi) after image acquisition at the end of each staining cycle. .. The following antibodies (all from Miltenyi Biotec) were used: Ki67, CD3, CD4, CD8, CD44, CD45RA, CD45RO, CD57, CD183, CD196, LAG3 (CD223), PD-1 (CD279), TIM3 (CD366), FoxP3, HLA-DR. Acquired images were processed and analyzed by MACSiQ ® View Imaging Software (Miltenyi Biotec) following current processing work-flow (Miltenyi Biotec). ..
Software:Article Title: Inhibition of autophagy enhances the antitumor efficacy of T/CAR T cell against neuroblastoma
Article Snippet: Fluorochromes were photobleached or released using REAlease technology (Miltenyi) after image acquisition at the end of each staining cycle. .. The following antibodies (all from Miltenyi Biotec) were used: Ki67, CD3, CD4, CD8, CD44, CD45RA, CD45RO, CD57, CD183, CD196, LAG3 (CD223), PD-1 (CD279), TIM3 (CD366), FoxP3, HLA-DR. Acquired images were processed and analyzed by MACSiQ ® View Imaging Software (Miltenyi Biotec) following current processing work-flow (Miltenyi Biotec). ..
Staining:Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells
Article Snippet: .. For phenotypic analysis, 0.2 × 10 6 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 10 6 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [ ] or the anti-La mAb (7B6) [ ] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 °C in the dark. .. Stained cells were analyzed by applying MACSQuant ® Analyzer 10 and MACSQuantify ® software (both from Miltenyi Biotec GmbH).
Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells.
Article Snippet: .. For phenotypic analysis, 0.2 × 106 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 106 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [45] or the anti-La mAb (7B6) [46] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 ◦C in the dark. .. Stained cells were analyzed by applying MACSQuant® Analyzer 10 and MACSQuantify® software (both from Miltenyi Biotec GmbH).
Expressing:Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells
Article Snippet: .. For phenotypic analysis, 0.2 × 10 6 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 10 6 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [ ] or the anti-La mAb (7B6) [ ] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 °C in the dark. .. Stained cells were analyzed by applying MACSQuant ® Analyzer 10 and MACSQuantify ® software (both from Miltenyi Biotec GmbH).
Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells.
Article Snippet: .. For phenotypic analysis, 0.2 × 106 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 106 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [45] or the anti-La mAb (7B6) [46] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 ◦C in the dark. .. Stained cells were analyzed by applying MACSQuant® Analyzer 10 and MACSQuantify® software (both from Miltenyi Biotec GmbH).
Construct:Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells
Article Snippet: .. For phenotypic analysis, 0.2 × 10 6 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 10 6 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [ ] or the anti-La mAb (7B6) [ ] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 °C in the dark. .. Stained cells were analyzed by applying MACSQuant ® Analyzer 10 and MACSQuantify ® software (both from Miltenyi Biotec GmbH).
Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells.
Article Snippet: .. For phenotypic analysis, 0.2 × 106 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 106 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [45] or the anti-La mAb (7B6) [46] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 ◦C in the dark. .. Stained cells were analyzed by applying MACSQuant® Analyzer 10 and MACSQuantify® software (both from Miltenyi Biotec GmbH).
Incubation:Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells
Article Snippet: .. For phenotypic analysis, 0.2 × 10 6 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 10 6 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [ ] or the anti-La mAb (7B6) [ ] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 °C in the dark. .. Stained cells were analyzed by applying MACSQuant ® Analyzer 10 and MACSQuantify ® software (both from Miltenyi Biotec GmbH).
Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells.
Article Snippet: .. For phenotypic analysis, 0.2 × 106 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 106 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [45] or the anti-La mAb (7B6) [46] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 ◦C in the dark. .. Stained cells were analyzed by applying MACSQuant® Analyzer 10 and MACSQuantify® software (both from Miltenyi Biotec GmbH).
Labeling:Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells
Article Snippet: .. For phenotypic analysis, 0.2 × 10 6 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 10 6 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [ ] or the anti-La mAb (7B6) [ ] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 °C in the dark. .. Stained cells were analyzed by applying MACSQuant ® Analyzer 10 and MACSQuantify ® software (both from Miltenyi Biotec GmbH).
Article Title: CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells.
Article Snippet: .. For phenotypic analysis, 0.2 × 106 cells were stained with monoclonal Abs (mAbs) against CD4, CD8, CD45RO, CD69, PD-1, Tim-3, Lag-3 (all purchased from Miltenyi Biotec GmbH), and CD62L (BioLegend, San Diego, CA, USA) for 15 min. To determine the percentage of cells expressing the respective RevCAR construct, 0.2 × 106 cells were incubated with 0.25 μg of the anti-La mAb (5B9) [45] or the anti-La mAb (7B6) [46] for 1 h, followed by staining with a fluorescently labeled secondary goat anti-mouse IgG Ab (BioLegend) for 30 min. All staining steps were performed at 4 ◦C in the dark. .. Stained cells were analyzed by applying MACSQuant® Analyzer 10 and MACSQuantify® software (both from Miltenyi Biotec GmbH).
|
Miltenyi Biotec is a verified supplier 
Miltenyi Biotec manufactures this product 
