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

Journal: bioRxiv

Article Title: A Cellular Cytotoxicity Assay using Ready-to-Thaw Target Cells without Washing Steps

doi: 10.64898/2026.01.21.700863

Figure Lengend Snippet: Toxicity of different concentrations of the cryopreservation medium STEM-CELLBANKER ® EX (SCB) towards K562 cells, primary NK cells, and NK-92 cells; cell viability was assessed via flow cytometry after 24 h incubation. A: Relative viability of K562 cells incubated with 0–100% SCB at 37°C and room temperature (RT) (n = 3 independent experiments, each 1 technical replicate); statistical analysis: two-way ANOVA with Tukey post hoc test within RT and 37°C groups. B: Relative viability of primary NK cells with 0–100% SCB at RT, 37°C, and 37°C + 500 U/mL IL-2 (left, n = 1 experiment, 1 technical replicate; no statistics performed) and with 0–25% SCB at 37°C (right, n = 2 donors/experiments, each 3 technical replicates); statistical analysis: two-way ANOVA with Tukey post hoc test. C: Relative viability of NK-92 cells with 0–100% SCB (left, n = 1 experiment, 3 technical replicates) and 0– 25% SCB (right, n = 1 experiment, 3 technical replicates) at 37°C; statistical analysis: one-way ANOVA with Tukey post hoc test. Bars represent mean ± SEM (ns = not significant; *p < 0.05; **p < 0.01; ****p < 0.0001).

Article Snippet: Purified cells were cultured in NK-MACS ® medium (Miltenyi Biotec, Bergisch Gladbach, Germany) with 5% human AB serum (PAN-Biotech), NK-MACS ® supplement (Miltenyi Biotec), 100 U/mL penicillin plus 100 μg/mL streptomycin (Sigma-Aldrich), and 500 U/mL recombinant human IL-2 IS (Miltenyi Biotec).

Techniques: Flow Cytometry, Incubation