Journal: bioRxiv
Article Title: TcrDesign: De novo design of epitope specific full-length T cell receptors
doi: 10.64898/2026.01.15.699824
Figure Lengend Snippet: ( A ) UMAP dimensionality reduction was performed on generated TCRs for two distinct epitopes, utilizing TCR embedding extracted by TcrDesign-B. ( B ) Binding assay of HLA-A*02:01-ELAGIGILTV tetramer to the nine generated TCRs expressing Jurkat cells was analyzed by flow cytometry. Three candidate TCRs ELA-TCR3, ELA-TCR4 and ELA-TCR6 exhibited specific binding to the tetramer. ( C ) Activation markers NFAT and CD69 levels of reporter Jurkat NFAT-ZsGreen cells expressing candidate TCRs ELA-TCR3, ELA-TCR4, ELA-TCR6 and ELApositive TCR after incubation with ELAGIGILTV-pulsed T2 cell for 24 hours, each sample was tested in triplicate. ( D ) The NFAT-ZsGreen signal and CD69 level of ELApositive-TCR and ELA-TCR3 were determined by fluorescence imaging and flow cytometry analysis. ( E ) Engineered Jurkat T cells were co-cultured with T2 cells and serially diluted peptides for 24 h. Co-cultured supernatants were analyzed by ELISA for secreted IL-2. ( F ) TCR-T cells at various E:T ratios were co-cultured with luciferase-transduced T2 cells. The % specific lysis of T2-luciferase (black) and 1ug/mL ELAGIGILTV pulsed T2-luciferase cells (blue) obtained by bioluminescence assay is plotted against multiple E:T ratios. Dots in the figure represents three replicates.
Article Snippet: For 1 x 10 6 T-lymphocytes, 25 μL of Dynabeads coated with anti-CD3 and anti-CD28 antibodies was added to the RPMI 1640 media supplemented with 200U human recombinant IL-2 (#11848-HNAH1-E, Sino Biological).
Techniques: Generated, Binding Assay, Expressing, Flow Cytometry, Activation Assay, Incubation, Fluorescence, Imaging, Cell Culture, Enzyme-linked Immunosorbent Assay, Luciferase, Lysis, ATP Bioluminescent Assay