rabbit antihuman 5t4 antibody (Danaher Inc)
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rabbit antihuman 5t4 antibody
Rabbit Antihuman 5t4 Antibody, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit antihuman 5t4 antibody/product/Danaher Inc
Average 86 stars, based on 1 article reviews
Rabbit Antihuman 5t4 Antibody, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit antihuman 5t4 antibody/product/Danaher Inc
Average 86 stars, based on 1 article reviews
rabbit antihuman 5t4 antibody - by Bioz Stars,
2026-03
86/100 stars
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1) Product Images from "Mechanistic and pharmacodynamic studies of DuoBody-CD3x5T4 in preclinical tumor models"
Article Title: Mechanistic and pharmacodynamic studies of DuoBody-CD3x5T4 in preclinical tumor models
Journal: Life Science Alliance
doi: 10.26508/lsa.202201481
Figure Legend Snippet: (A) TPBG (5T4) mRNA expression from the The Cancer Genome Atlas expression database across different tumor indications. Data are shown with each single dot representing a single tumor and with the mean fragments per kilobase million ± SD for each indication. (B, C) 5T4 protein expression was evaluated by IHC in tumor TMAs. (B) One representative tissue core is shown from each of the evaluated cancer indications. In the top row, the scale bar indicates either 500 μm (breast, bladder, uterine, lung, and esophageal cancer) or 1 mm (HNSCC and prostate), whereas in the bottom row, the scale bar corresponds to 50 μm. (C) The percentage of 5T4 + tumor cells in each tumor type, as well as the median ± min/max values (error bars) and the quartiles (box) for each indication as determined by IHC analysis. The gray shading represents the cutoff (≥10% of tumor cells that are 5T4 + ) used to determine the prevalence of 5T4 expression (Table S1).
Techniques Used: Expressing
Figure Legend Snippet: (A) Binding of DuoBody-CD3x5T4 to recombinant 5T4 as determined by BLI. (B) CHO-S cells, transiently transfected with human 5T4, were stained with DuoBody-CD3x5T4 or negative control antibodies bsIgG1-CD3xctrl and IgG1-ctrl-K409R. Non-transfected CHO-S cells were included as negative control for binding of DuoBody-CD3x5T4. Data shown are the geometric mean fluorescence intensity ± SD of duplicate wells from one representative flow cytometry experiment of three experiments performed. (C) Binding of DuoBody-CD3x5T4 to recombinant CD3ε as determined by BLI. (D) Jurkat T cells were stained with DuoBody-CD3x5T4. As a negative control, IgG1-ctrl-K409R (30 μg/ml) was included. Data shown are geometric mean fluorescence intensity ± SD of duplicate wells from one representative experiment of three flow cytometry experiments performed.
Techniques Used: Binding Assay, Recombinant, Transfection, Staining, Negative Control, Fluorescence, Flow Cytometry
Figure Legend Snippet: (A) Schematic representation of the T cell–mediated cytotoxicity assay and its readouts. Cocultures were incubated for 72 h at an E:T ratio of 4:1, unless specified otherwise. (B, C) MDA-MB-468 tumor cells (∼32,000 5T4 molecules/cell as determined by quantitative flow cytometry) were incubated with purified T cells from three different donors at the indicated E:T ratios and DuoBody-CD3x5T4 or the control antibodies bsIgG1-ctrlx5T4 or bsIgG1-CD3xctrl for 72 h. (B) Data shown are mean percentage survival ± SD of duplicate wells from one representative donor of three donors included in three experiments. (C) Median IC 50 (±range) of loss of tumor cell viability for all tested E:T ratios. (D, E, F, G, H) T cell–mediated cytotoxicity assay with MDA-MB-231 tumor cells (∼14,000 5T4 molecules/cell as determined by quantitative flow cytometry) was performed with purified T cells (E:T ratio = 4:1, n = 3 donors) and DuoBody-CD3x5T4 for 24, 48, and 72 h. Shown is a representative donor of three donors tested. (D) Mean percentages of viable tumor cells ± SD of duplicate wells is shown, illustrating the kinetics of DuoBody-CD3x5T4–induced T cell–mediated cytotoxicity of MDA-MB-231 cells. (E) Kinetics of CD4 + (left) and CD8 + (right) T-cell activation (CD69 expression) induced by DuoBody-CD3x5T4 when incubated with MDA-MB-231 cells. (F) Kinetics of IFNγ, IL-6, IL-8, and TNFα production induced by DuoBody-CD3x5T4 when cocultured with T cells and MDA-MB-231 cells, showing a representative donor of two donors tested. (G, H) Kinetics of granzyme B (G) and perforin (H) production induced by DuoBody-CD3x5T4 when cocultured with T cells and MDA-MB-231 cells, showing a representative donor of two donors tested. (I) MDA-MB-231 tumor cells were incubated with CFSE-labeled T cells (E:T ratio = 8:1) and DuoBody-CD3x5T4 or control antibodies for 72 h. CFSE dilution in T cells was analyzed by flow cytometry and the T-cell expansion index (i.e., how much the total T-cell population has expanded by proliferation) for CD4 + (left panel) and CD8 + (right panel) T cells is shown. Data shown are a representative donor of three donors tested.
Techniques Used: Cytotoxicity Assay, Incubation, Flow Cytometry, Purification, Control, Activation Assay, Expressing, Labeling
Figure Legend Snippet: (A) Expression of 5T4 in a panel of cancer cell lines of different indications, determined by quantitative flow cytometry (n = 2–4 per cell line). Shown here are the mean (horizontal line) and the range of expression detected. (B, C, D) Dose-dependent T cell–mediated cytotoxicity of different tumor cell lines in the presence of DuoBody-CD3x5T4 and purified T cells (E:T ratio = 4:1) after 72 h. Shown are mean survival percentages ± SD of duplicate wells derived from a representative donor of five (RL95-2) or three (SW780, SK-GT-4) donors tested. (E) Geomean IC 50 values (and range) for DuoBody-CD3x5T4–induced loss of tumor cell viability of all tested tumor cell lines (n = 3–10 donors/cell line) plotted against the number of 5T4 molecules/cell ( P ≤ 0.05; nonparametric Spearman correlation). (F) T cell–mediated cytotoxicity of MDA-MB-231 parental and 5T4 KO cells in the presence of DuoBody-CD3x5T4 and purified T cells (E:T ratio = 4:1) after 72 h, showing a representative donor of 4 donors tested.
Techniques Used: Expressing, Flow Cytometry, Purification, Derivative Assay
Figure Legend Snippet: (A, B, C, D, E, F, G, H, I, J, K, L) T cell–mediated cytotoxicity induced by DuoBody-CD3x5T4 when incubated with purified T cells from one representative donor (E:T ratio = 4:1) and a range of cancer cell lines with varying levels of 5T4 expression for 72 h: (A) SiHa (n = 6 donors tested), (B) NCI-H292 (n = 5 donors tested), (C) BxPc-3 (n = 3 donors tested), (D) MDA-MB-231 (n = 5 donors tested), (E) PANC-1 (n = 9 donors tested), (F) Fadu (n = 4 donors tested), (G) EPLC-272H (n = 3 donors tested), (H) Ca Ski (n = 5 donors tested), (I) RT-122 (n = 6 donors tested), (J) PC-3 (n = 3 donors tested), (K) SCC-9 (n = 4 donors tested), and (L) DU 145 (n = 5 donors tested). (M) 5T4 expression on MDA-MB-231 parental and 5T4 KO cells was measured by flow cytometry.
Techniques Used: Incubation, Purification, Expressing, Flow Cytometry
Figure Legend Snippet: (A) 5T4 and Fas expression on MDA-MB-231 parental and Fas KO cells was measured by quantitative flow cytometry with or without prior overnight incubation with IFNγ (100 ng/ml). (B, C, D, E) MDA-MB-231 parental and Fas KO cells were incubated with purified CD4 + (B, D) or CD8 + (C, E) T cells (E:T ratio = 4:1, n = 5 donors) and DuoBody-CD3x5T4 for 72 h. T-cell activation (B, C) and T cell–mediated cytotoxicity (D, E) were analyzed. (B, C) Activation of T cells from five donors at 1 μg/ml DuoBody-CD3x5T4 (* P ≤ 0.05, paired t test). (D, E) The left panels show dose-dependent T cell–mediated cytotoxicity in a representative experiment, and the right panels show tumor cell viability at 1 μg/ml DuoBody-CD3x5T4 from five donors (* P ≤ 0.05 and ** P ≤ 0.01, paired t test). (F) 5T4, IFNGR1 and Fas expression on MDA-MB-231 parental and IFNGR1 KO cells was measured by flow cytometry. (G, H) MDA-MB-231 parental and IFNGR1 KO cells were incubated with purified T cells (E:T ratio = 4:1, n = 4 donors) and DuoBody-CD3x5T4 for 72 h. (G) The left panel shows dose-dependent T cell–mediated cytotoxicity in a representative experiment, and the right panel shows tumor cell viability at 1 μg/ml DuoBody-CD3x5T4 from four donors (*** P ≤ 0.005, paired t test). (H) T-cell activation at 1 μg/ml DuoBody-CD3x5T4 from four donors.
Techniques Used: Expressing, Flow Cytometry, Incubation, Purification, Activation Assay
Figure Legend Snippet: (A) MDA-MB-231 parental and Fas KO cells were incubated with purified CD4 + or CD8 + T cells (E:T ratio = 4:1, n = 5 donors) and DuoBody-CD3x5T4 for 72 h. T-cell activation was analyzed by measuring up-regulation of CD69. (B) 5T4 and Fas expression on NCI-H1299 parental and Fas KO cells was measured by flow cytometry with or without incubation with IFNγ (100 ng/ml). (C, D) NCI-H1299 parental and Fas KO cells were incubated with purified CD4 + (top panels) or CD8 + (bottom panels) T cells (E:T ratio = 4:1, n = 5–6 donors tested) and DuoBody-CD3x5T4 for 72 h. (C) The left panels show dose-dependent T cell–mediated cytotoxicity, and the right panels show T cell–mediated cytotoxicity at 1 μg/ml DuoBody-CD3x5T4. A paired t test was used to compare T cell–mediated cytotoxicity at 1 μg/ml, with * P ≤ 0.05 and ** P ≤ 0.01. (D) The left panels show dose-dependent T-cell activation, and the right panels show T-cell activation at 5 μg/ml DuoBody-CD3x5T4. (E) MDA-MB-231 parental and IFNGR1 KO cells were incubated with purified T cells (E:T ratio = 4:1, n = 4 donors) and DuoBody-CD3x5T4 for 72 h, and T-cell activation was analyzed measuring up-regulation of CD69. A representative donor of four donors tested is shown.
Techniques Used: Incubation, Purification, Activation Assay, Expressing, Flow Cytometry
Figure Legend Snippet: (A) Parental and 5T4 KO MDA-MB-231 tumor cells were mixed in different ratios, as indicated, and incubated with purified T cells (E:T ratio = 4:1, n = 2 donors) and DuoBody-CD3x5T4 for 72 h. T cell–mediated cytotoxicity of 5T4 + (left panel) and 5T4 − (right panel) tumor cells was determined by flow cytometry, showing a representative donor of three donors tested. (B, C, D) Parental (5T4 + ) MDA-MB-231 tumor cells were cocultured with purified T cells (E:T = 4:1, n = 2 donors) and incubated with 10 μg/ml DuoBody-CD3x5T4 or bsIgG1-CD3xctrl for 72 h. As a positive control, T cells were incubated with anti-CD3/CD28 beads (but without tumor cells) for 72 h. (B) The supernatant (either with or without T cells) was transferred to MDA-MB-231 5T4 KO cells and incubated for 72 h (B). As negative control, MDA-MB-231 5T4 KO cells were incubated with fresh T cells and indicated antibodies for 72 h. (C, D) T cell–mediated cytotoxicity (C) and Fas expression (D) of MDA-MB-231 5T4 KO cells were determined by flow cytometry. A representative donor of two donors tested is shown. (E) 5T4, Fas, IFNGR1, and PD-L1 expression on MDA-MB-231 parental, 5T4/Fas KO, and 5T4/IFNGR1 KO cells was measured by flow cytometry with or without prior overnight incubation with IFNγ (100 ng/ml). (F, G, H, I) Parental and 5T4/Fas KO or 5T4/IFNGR1 MDA-MB-231 tumor cells were mixed in different ratios, as indicated, and incubated with purified T cells (E:T ratio = 4:1, n = 4–6) and 1 μg/ml DuoBody-CD3x5T4 for 72 h, after which T cell–mediated cytotoxicity (F), CD4 + (G) and CD8 + (H) T-cell activation, and IFNγ production (I) were analyzed. The predicted black line in (F) refers to the outcome of the assay when no bystander killing is expected, for example, if 50% of tumor cells are 5T4 + , only 50% of tumor cells will be killed.
Techniques Used: Incubation, Purification, Flow Cytometry, Positive Control, Negative Control, Expressing, Activation Assay
![(A, B) Parental (5T4 + ) MDA-MB-231 tumor cells were cocultured with purified T cells (E:T = 4:1, n = 2 donors) and incubated with 10 μg/ml DuoBody-CD3x5T4 or bsIgG1-CD3xctrl for 72 h. As a positive control, T cells were incubated with anti-CD3/CD28 beads (but without tumor cells) for 72 h. The supernatant (either with or without T cells) was transferred to MDA-MB-231 5T4 KO cells and incubated for 72 h. As negative control, MDA-MB-231 5T4 KO cells were incubated with fresh T cells and indicated antibodies for 72 h. (A, B) T cell–mediated cytotoxicity of MDA-MB-231 parental cells (A) and T-cell activation (B) were determined by flow cytometry. (C, D, E) Parental and 5T4, 5T4/Fas, or 5T4/IFNGR1 KO MDA-MB-231 tumor cells were mixed in different ratios, as indicated, and incubated with purified T cells (E:T ratio = 4:1) and DuoBody-CD3x5T4 for 72 h, after which T cell–mediated cytotoxicity (C, three representative donors of 4 [5T4/IFNGR1 KO] or six [5T4 and 5T4/Fas KO] analyzed), CD8 + T-cell activation (D, a representative donor of 4 [5T4/IFNGR1 KO] or six [5T4 and 5T4/Fas KO] analyzed), and IFNγ production (E, one representative donor of 4 [5T4/IFNGR1 KO] or six [5T4 and 5T4/Fas KO] analyzed) were determined.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_8754/pmc09458754/pmc09458754__LSA-2022-01481_FigS7.jpg "(A, B) Parental (5T4 + ) MDA-MB-231 tumor cells were cocultured with ...")
Figure Legend Snippet: (A, B) Parental (5T4 + ) MDA-MB-231 tumor cells were cocultured with purified T cells (E:T = 4:1, n = 2 donors) and incubated with 10 μg/ml DuoBody-CD3x5T4 or bsIgG1-CD3xctrl for 72 h. As a positive control, T cells were incubated with anti-CD3/CD28 beads (but without tumor cells) for 72 h. The supernatant (either with or without T cells) was transferred to MDA-MB-231 5T4 KO cells and incubated for 72 h. As negative control, MDA-MB-231 5T4 KO cells were incubated with fresh T cells and indicated antibodies for 72 h. (A, B) T cell–mediated cytotoxicity of MDA-MB-231 parental cells (A) and T-cell activation (B) were determined by flow cytometry. (C, D, E) Parental and 5T4, 5T4/Fas, or 5T4/IFNGR1 KO MDA-MB-231 tumor cells were mixed in different ratios, as indicated, and incubated with purified T cells (E:T ratio = 4:1) and DuoBody-CD3x5T4 for 72 h, after which T cell–mediated cytotoxicity (C, three representative donors of 4 [5T4/IFNGR1 KO] or six [5T4 and 5T4/Fas KO] analyzed), CD8 + T-cell activation (D, a representative donor of 4 [5T4/IFNGR1 KO] or six [5T4 and 5T4/Fas KO] analyzed), and IFNγ production (E, one representative donor of 4 [5T4/IFNGR1 KO] or six [5T4 and 5T4/Fas KO] analyzed) were determined.
Techniques Used: Purification, Incubation, Positive Control, Negative Control, Activation Assay, Flow Cytometry
Figure Legend Snippet: (A) Experimental outline of the evaluation of T cell–mediated tumor cell killing by DuoBody-CD3x5T4 in dissociated patient-derived solid tumor samples ex vivo. (B) 5T4 and CD3 expression in ovarian tumor samples OV1 and OV2, as determined by IHC. The scale bars in the top row correspond to 500 μm, and the scale bars in the bottom row correspond to 50 μm (OV1) or 20 μm (OV2). (C) Binding of DuoBody-CD3x5T4 or isotype control (bsIgG1-CD3xctrl) to dissociated patient-derived solid tumor cells (CD45 − population) was evaluated by flow cytometry. LU, lung cancer; OV, ovarian cancer; UT, uterine cancer. (D) The percentage of CD3 + T cells in the dissociated patient-derived solid tumor samples. (E, F, G, H) T cell–mediated cytotoxicity ± SEM (E), T-cell activation (F; %CD25 + and PD-1 + ), and cytokine (G) and GZMB (H) production ± SD of duplicate wells induced by DuoBody-CD3x5T4 versus control (bsIgG1-CD3xctrl) in four dissociated patient-derived solid tumor samples ex vivo.
Techniques Used: Derivative Assay, Ex Vivo, Expressing, Binding Assay, Control, Flow Cytometry, Activation Assay