simple plex human total tau  (Protein Simple Inc)

Journal: Journal of Translational Medicine

Article Title: Multi-omic profiling and preclinical efficacy of fratricide-driven, unedited CD7 CAR-T cells in T-cell leukemia

doi: 10.1186/s12967-026-07701-5

Figure Lengend Snippet: Tech transfer of UMCG-001 manufacturing to GMP-facility. ( A ) Overview for three UMCG-001 batches produced in a CliniMACS Prodigy ® bioreactor. Batch 1 and 3 were manufactured for 6 days, with cell harvest on day 6. Culture of batch 2 was expanded to day 12. For all, cell selection and activation were performed on day 0, LV transduction on day 1, HPL supplementation on day 4 and quality and characterization tests on harvesting day. Parallel lab batches were generated with matched-donor material for batch 2 and 3. ( B ) Flow cytometry-based quantification of transduction efficiency as %GFP + cells. ( C ) UMCG-001 expansion in million cells over time. Dotted line: DP release specification. ( D ) Fold-change expansion exhibited by UMCG-001 batch 3 generated at the GMP facility or Lab. ( E ) Fold-change expansion exhibited by UMCG-001 batch 2 generated at the GMP facility or Lab. ( F ) Flow cytometry scatterplots illustrating cell viability on harvesting day assessed by LIVE/DEAD TM staining within T-cell gate. ( G ) Histogram representation of CD7 expression on manufacturing day 6 among the different batches and CD19 CAR-T cells as reference. ( H ) Flow cytometry scatterplots representing CD7 vs. K12 CAR (GFP + ) expression across the batches on day 6. ( I ) Killing capacity across UMCG-001 batches of CD7 + and CD7 Neg cell lines at different E: T ratios for 24 h as AnnexinV/PI flow cytometry-based quantification. ( J ) Specific OV-CAR-3.Luc.CD7 killing calculated as indicated in Formula 1 by GMP batch 1 up to 5 rounds of 24 h-tumor re-challenges at different E: T ratios or ( K ) Percentage of viable cells at selected 2:1, 1:1 and 1:4 E: T. ( L ) IFN-γ secretion by the GMP batches when co-cultured with non-loaded (NL), CD7-loaded (CD7L), CD19-loaded (CD19L) or no beads for 24 h on day 6. Delta IFN-γ secretion in respect to NL beads. ( M ) IFN-γ secretion by UMCG-001 batches vs. CD19 CAR-T cells batches or ( N ) matched GMP and lab batch 3 as in ( L )

Article Snippet: IFN-γ secretion was quantified using the Simple Plex Human IFN-γ (3rd Gen) Cartridge on the ELLA platform (Protein Simple, Bio-Techne, Minneapolis, USA) as previously described [ ].

Techniques: Produced, Selection, Activation Assay, Transduction, Generated, Flow Cytometry, Staining, Expressing, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Multi-omic profiling and preclinical efficacy of fratricide-driven, unedited CD7 CAR-T cells in T-cell leukemia

doi: 10.1186/s12967-026-07701-5

Figure Lengend Snippet: Autologous manufacturing and malignant contamination assessment in UMCG-001. ( A ) UMCG-001 manufactured from T-ALL patient 1 (T-ALL_1) derived material. ( B ) IFN-γ secretion upon 72 h co-culture of UMCG-001 batches with CD7 + (Jurkat, MOLT-4), autologous T-ALL and CD7 Neg K-562 cells. ( C ) Flow cytometry-based assessment of the percentage of killing of CD7 + (Jurkat, MOLT-4), autologous T-ALL and CD7 Neg K-562cells by UMCG-001. ( D ) Flow cytometry dot blot illustrating CD7 and surface CD3 (sCD3) expression in UMCG-001 and CD19 CAR-T cells upon fratricide. ( E ) Percentage of surface CD7 and CD3 over time in T.ALL_1 UMCG-001 final DP. Day − 1 represents the day before activation, day 0 after activation and transduction, and day 16 end of the fratricidal phase. ( F ) Schematic representation of sample preparation for subsequent single-cell RNA sequencing. T-ALL_1 original sample was composed of healthy (sCD3 + ) and blasts (sCD3 - ), which were sorted based on sCD3 expression. The whole sample was subjected to CAR - T manufacturing, after which CAR + cells were sorted based on GFP expression. All sorted samples were processed for V( D )J and gene expression (GEX) single-cell libraries. ( G ) Violin plot representation of genes related to malignant phenotype across TALL_1 sample. ( H ) Relative abundance of TCR clones across CD19 CAR-T cells UMCG-001 and sCD3 + samples, calculated as the number of unique TCRs per total number of cells. ( I ) Clone sizes across CD19 CAR-T cells and UMCG-001 batches generated from independent healthy donors (gray) and T-ALL_1 (black). The number of cells per clone group is shown

Article Snippet: IFN-γ secretion was quantified using the Simple Plex Human IFN-γ (3rd Gen) Cartridge on the ELLA platform (Protein Simple, Bio-Techne, Minneapolis, USA) as previously described [ ].

Techniques: Derivative Assay, Co-Culture Assay, Flow Cytometry, Dot Blot, Expressing, Activation Assay, Transduction, Sample Prep, Single Cell, RNA Sequencing, Gene Expression, Clone Assay, Generated