Journal: Science Advances

Article Title: Discovery and preclinical evaluation of monoclonal antibodies and bispecific engagers targeting the NKG2A inhibitory receptor

doi: 10.1126/sciadv.adu0690

Figure Lengend Snippet: ( A ) The IFN-⍺ + , TNFα + , and GrzB + population (%) of IL-2–activated NK cells after antibodies treatment with different NKG2A binding valency (monovalent, bivalent, tetravalent, or multivalent) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Internalization of NKG2A antibodies in NK cells, assessed by labeling the antibody with a pH-sensitive dye to monitor endocytosis. Durvalumab and pertuzumab (ptz IgG1) were included as controls. (A and B) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Primary NK cells were cultured with magnetic-activated cell sorting (MACS) basal NK media with supplementary (Miltenyi Biotec, 130-114-429), 10% (v/v) human serum (Sigma-Aldrich), and human IL-2 (hIL-2) (50 IU/ml; Miltenyi Biotec).

Techniques: Binding Assay, Labeling

Journal: Science Advances

Article Title: Discovery and preclinical evaluation of monoclonal antibodies and bispecific engagers targeting the NKG2A inhibitory receptor

doi: 10.1126/sciadv.adu0690

Figure Lengend Snippet: ( A ) CD16 + , NKG2A + , or NKG2C + population (%) of CD56 + NK cells after antibodies (100 nM) and IL-2 treatment (50 IU/ml) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Left: Graphs showing the cell killing activity of aHER2 × aNKG2A BiNK cotreatment with anti-HER2 IgG1 pertuzumab (ptz IgG1) in A549 and H2030 cells in the presence of primary NK cells. E:T ratio of 2:1. Right: Schematic figure of antibodies treatment schedule. (A) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05 and ** P < 0.01.

Article Snippet: Primary NK cells were cultured with magnetic-activated cell sorting (MACS) basal NK media with supplementary (Miltenyi Biotec, 130-114-429), 10% (v/v) human serum (Sigma-Aldrich), and human IL-2 (hIL-2) (50 IU/ml; Miltenyi Biotec).

Techniques: Activity 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