human il 2 research grade  (Miltenyi Biotec)

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: Nanobody MET CAR T cells show efficacy in solid tumors

doi: 10.64898/2026.01.27.702111

Figure Lengend Snippet: (A) Expression level of mRNA VHH2-BBz and VHH2-28z in primary human T cells. (B) Avidity assay using Lumicks z-Movi, comparison between VHH2-BBz and VHH2-28z (n = 3 technical replicates; two-way ANOVA). (C) Comparison of MDA-MB-231 killing between VHH2-BBz and VHH2-28z (n=8 technical replicates, two-way ANOVA). (D) Intracellular cytokine production analyzed by flow cytometry; cells harvested 16 hours after co-culture with MDA-MB-231 in the presence of 2μM of the Golgi inhibitor monensin (n = 4 technical replicates, two-way ANOVA). (E) Polyfunctioinal strength index (PSI) using IsoPlexis, comparison between CD8+ VHH2-BBz and CD8+ VHH2-28z 16 hours after co-culture with Karpas-422. Effector cytokines (Granzyme B, IFN-γ, MIP-1a, perforin, TNF-α, TNF-β); Stimulatory cytokines (GM-CSF, IL-12, IL-15, IL-2, IL-21, IL-5, IL-7, IL-8, IL-9); Chemoattractive cytokines (CCL-11, IP-10, MIP-1b, RANTES); Regulatory cytokines (IL-10, IL-13, IL-22, IL-4, sCD137, sCD40L, TGF-b1); Inflammatory cytokines (IL-17A, IL-17F, IL-1b, IL-6, MCP-1, MCP-4). (F-I) Microwell assay using TROVO, comparing single cell killing rate (F) (n=3 technical replicates; two-tailed Student’s t -test), T cell proliferation (G) (n=2 donorsx3 replicates; two-tailed Student’s t -test), and tumor killing (H) (n=2 donors x 3 replicates each, two-tailed Student’s t -test). (I) Representative images used to calculate data shown in (G) and (H). Each row represents one microwell monitored over 4 days. Each microwell is set up with E:T =20:10. T cell = red. * P<0.05; **P<0.01; ***P:<0.001; ****P:<0.0001 .

Article Snippet: Bulk T cells were isolated using the EasySep Human T Cell Enrichment Kit (STEMCELL Technologies, Cat. #17951), activated with ImmunoCult Human CD3/CD2/CD28 T cell activator (STEMCELL, Cat. #10990) at the recommended concentration, and cultured at 1 × 10 6 cells per mL in RPMI1640 + 10% FBS in the presence of 100 IU/mL recombinant human IL-2 (Miltenyi Biotec, Cat. #130-097743).

Techniques: Expressing, Comparison, Flow Cytometry, Co-Culture Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Nanobody MET CAR T cells show efficacy in solid tumors

doi: 10.64898/2026.01.27.702111

Figure Lengend Snippet: (A) Different binding epitopes predicted by AlphaFold3 for VHH2, scFv1, and scFv2. (B) Expression of compared CARs. (C) Avidity measurement by Lumicks z-Movi using MDA-MB-231 as target cells. N = 3 technical replicates, with P values from two-tailed Student’s t- test. (D) Avidity measurement against the 786-O cell line. N = 4-5 technical replicates; P values from two-tailed Student’s t- test. (E) Intracellular cytokine production by flow cytometry. Cells were stained before and 16-hours after stimulation by Karpas-422. N=3 technical replicates; P values from two-way ANOVA. (F) Polyfunctional strength index by IsoPlexis. Effector cytokines (Granzyme B, IFN-γ, MIP-1a, perforin, TNF-α, TNF-β); Stimulatory cytokines (GM-CSF, IL-12, IL-15, IL-2, IL-21, IL-5, IL-7, IL-8, IL-9); Chemoattractive cytokines (CCL-11, IP-10, MIP-1b, RANTES); Regulatory cytokines (IL-10, IL-13, IL-22, IL-4, sCD137, sCD40L, TGF-b1); Inflammatory cytokines (IL-17A, IL-17F, IL-1b, IL-6, MCP-1, MCP-4). (G-I) Hydrogel-based microwell co-culture assay by TROVO, showing single cell killing rate (G, 1:10 E:T), killing (H, 20:10 E:T), and T cell proliferation (I, 20:10 E:T). N=3 technical replicates; P values using two-tailed Student’s t- test. (J) Representative images of microwell co-culture at 20:10 E:T. Each row represents one microwell monitored over time. * P<0.05; **P<0.01; ***P<0.001; ****P<0.0001 .

Article Snippet: Bulk T cells were isolated using the EasySep Human T Cell Enrichment Kit (STEMCELL Technologies, Cat. #17951), activated with ImmunoCult Human CD3/CD2/CD28 T cell activator (STEMCELL, Cat. #10990) at the recommended concentration, and cultured at 1 × 10 6 cells per mL in RPMI1640 + 10% FBS in the presence of 100 IU/mL recombinant human IL-2 (Miltenyi Biotec, Cat. #130-097743).

Techniques: Binding Assay, Expressing, Two Tailed Test, Flow Cytometry, Staining, Co-culture Assay, Co-Culture 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