recombinant human ceacam5 fc  (Sino Biological)

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: Design and functional evaluation of ATOR-4066. A, Structure of ATOR-4066 generated in the bispecific RUBY format. B, Mode of action of ATOR-4066, as proposed by Hägerbrand and colleagues . C, The ability of serially diluted ATOR-4066 to induce CEACAM5 conditional activation of primary human MoDCs when cocultured for 48 hours with a sublethally UV-irradiated human CEACAM5-expressing cell line (MKN45) was assessed by measuring IL12 secretion in the supernatant by ELISA. Mean ± SD of normalized IL12 levels of nine donors was calculated, and EC 50 values were determined. D, The ability of ATOR-4066 to induce CEACAM5-conditional CD40 agonistic effects in a CD40 reporter assay was assessed in the presence of soluble CEACAM5 compared with a control CD40xCEACAM5 bsAb #3 with a high-affinity CEACAM5 binder targeting the same epitope as ATOR-4066, demonstrating that the CEACAM5 affinity is important. E, DTCs, including tumor-infiltrating CD40 + immune cells and CEACAM5 + tumor cells, obtained from patients with gastric cancer ( n = 6), were stimulated with ATOR-4066 in vitro for 72 hours, and activation markers, CD83 on myeloid cells and B cells, and CD25 on CD4 and CD8 T cells, were assessed using flow cytometry. F, Harvest titer of ATOR-4066 following production in 0.5 L and 1 L shaker flasks and in 2 L bioreactor. *, P ≤ 0.05; ns, not significant.

Article Snippet: ATOR-4066 or the control CD40-CEACAM5 bsAb #3 was coupled to anti-Human IgG Fc Capture biosensor tips (Sartorius) and assayed against 1:2 serially diluted monomeric CEACAM5 (4128-CM, R&D Systems), starting at 100 nmol/L.

Techniques: Functional Assay, Generated, Activation Assay, Irradiation, Expressing, Enzyme-linked Immunosorbent Assay, Reporter Assay, Control, In Vitro, Flow Cytometry

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: ATOR-4066 demonstrates strong antitumor activity in syngeneic MC38-expressing human CEACAM5. A, The dose-dependent response of ATOR-4066 was studied in F1 hCD40tgxC57BL/6 mice inoculated with MC38-CEA and treated with the indicated dose of ATOR-4066 or vehicle 6, 10, and 13 days after inoculation, n = 10–27. B, The antitumor effect of ATOR-4066 was compared with an equimolar dose of CD40 mAb and vehicle. C, Flow cytometry analysis and IHC were used to determine CEACAM5 expression in tumors at the indicated time points after cell inoculation in untreated mice, n = 6–10. D, The IHC score was used to determine antibody accumulation in mice treated with 167 μg ATOR-4066, 100 μg of CD40 mAb IHC, or isotype control. Bars represent mean ± SD; each point represents one mouse; n = 5. E and F, Kaplan–Meier plots of mice in which ATOR-4066 treatment (167 µg/dose) was started either 10 days after inoculation ( E ) or 13 days after inoculation ( F ); n = 20. Data represent a representative experiment (from two independent experiments) performed ( B and D ) or pooled data from two ( C , E , F ) independent experiments performed. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. MFI, mean fluorescence intensity.

Article Snippet: ATOR-4066 or the control CD40-CEACAM5 bsAb #3 was coupled to anti-Human IgG Fc Capture biosensor tips (Sartorius) and assayed against 1:2 serially diluted monomeric CEACAM5 (4128-CM, R&D Systems), starting at 100 nmol/L.

Techniques: Activity Assay, Expressing, Flow Cytometry, Control, Fluorescence

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: ATOR-4066 alters the transcription profiles of tumors and intratumoral immune cells to a proinflammatory state. A, Transcriptomic alterations induced by ATOR-4066 in blood, tumors, and intratumoral immune cells were studied in F1 hCD40tgxC57BL/6 mice inoculated with MC38-CEACAM5 tumor cells. B and C, PCA illustrated large transcriptomic changes in tumors but not in peripheral blood following ATOR-4066 treatment. D, Gene expression profiles of all DEGs in tumors between the treatment conditions were clustered, and GSEA was performed on each gene cluster. E, GSVA was performed on the top enriched and CD40-related pathways. F, Gene expression of selected canonical inflammatory or immune activation genes. G, The activity of transcription factors (TF) was estimated, and the activity of the top varying transcription factors is shown. H, Gene expression profiles of all DEGs in intratumoral immune cells between the treatment conditions were clustered, and GSEA was performed on each gene cluster. I and J, Immune cell composition was estimated based on CIBERSORTx, and canonical genes for cell populations of immune activation were analyzed. *, P ≤ 0.05; **, P ≤ 0.01. NA, not applicable; Pos., positive; Reg., regulation; VST, variance stabilization transformed.

Article Snippet: ATOR-4066 or the control CD40-CEACAM5 bsAb #3 was coupled to anti-Human IgG Fc Capture biosensor tips (Sartorius) and assayed against 1:2 serially diluted monomeric CEACAM5 (4128-CM, R&D Systems), starting at 100 nmol/L.

Techniques: Gene Expression, Activation Assay, Activity Assay, Transformation Assay

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: ATOR-4066 in combination with the PD-1 inhibitor enhances the activation of exhausted T cells and controls tumor growth. A and B, ATOR-4066 was evaluated in combination with the anti–PD-1 antibody Opdivo in MLR assays using allogeneic exhausted human primary CD4 + or CD8 + T cells and mature MoDCs. CD4 + or CD8 + T cells with an exhausted phenotype were expanded for 7 days with Dynabeads and then rested overnight in medium. Titrations of either ATOR-4066 and a constant concentration of anti–PD-1 or titrations of anti–PD-1 and a constant concentration of ATOR-4066 in the presence of CEACAM5 beads for cross-linking were used in cultures with a 1:10 mix of mature MoDC cells and exhausted T cells for 7 days before cell culture supernatants were harvested for analyzing IFNγ levels by ELISA. A, Dose–response curves (normalized data, background subtracted, and mean and SD from 9, 14, and 11 donors) are displayed. B, T-cell activation of exhausted CD8 T cells was higher for the combination of ATOR-4066 and PD-1 compared with the sum of the monospecific activity at higher concentrations, exemplified at 1 nmol/L ATOR-4066 and 100 nmol/L anti–PD-1. C and D, To study the in vivo combination treatment effect, F1 hCD40tgxC57BL/6 mice were inoculated with MC38-CEA and treated with 50 μg anti–PD-1 on days 10 and 13 after inoculation and/or 60 μg ATOR-4066 on days 6, 10, and 13 after inoculation. Survival was recorded, and tumor volume was measured. Data represent pooled data from two experiments independently performed. n = 18–20/group. conc., concentration.

Article Snippet: ATOR-4066 or the control CD40-CEACAM5 bsAb #3 was coupled to anti-Human IgG Fc Capture biosensor tips (Sartorius) and assayed against 1:2 serially diluted monomeric CEACAM5 (4128-CM, R&D Systems), starting at 100 nmol/L.

Techniques: Activation Assay, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Activity Assay, In Vivo

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: Design and functional evaluation of ATOR-4066. A, Structure of ATOR-4066 generated in the bispecific RUBY format. B, Mode of action of ATOR-4066, as proposed by Hägerbrand and colleagues . C, The ability of serially diluted ATOR-4066 to induce CEACAM5 conditional activation of primary human MoDCs when cocultured for 48 hours with a sublethally UV-irradiated human CEACAM5-expressing cell line (MKN45) was assessed by measuring IL12 secretion in the supernatant by ELISA. Mean ± SD of normalized IL12 levels of nine donors was calculated, and EC 50 values were determined. D, The ability of ATOR-4066 to induce CEACAM5-conditional CD40 agonistic effects in a CD40 reporter assay was assessed in the presence of soluble CEACAM5 compared with a control CD40xCEACAM5 bsAb #3 with a high-affinity CEACAM5 binder targeting the same epitope as ATOR-4066, demonstrating that the CEACAM5 affinity is important. E, DTCs, including tumor-infiltrating CD40 + immune cells and CEACAM5 + tumor cells, obtained from patients with gastric cancer ( n = 6), were stimulated with ATOR-4066 in vitro for 72 hours, and activation markers, CD83 on myeloid cells and B cells, and CD25 on CD4 and CD8 T cells, were assessed using flow cytometry. F, Harvest titer of ATOR-4066 following production in 0.5 L and 1 L shaker flasks and in 2 L bioreactor. *, P ≤ 0.05; ns, not significant.

Article Snippet: In brief, CD40 reporter cells and ATOR-4066 or control bsAb (CD40xCEA bsAb #3) were added to the assay plates before the addition of CEACAM5-transfected CHO cells in the presence or absence of soluble CEACAM5 (0.5, 1, or 5 μg/mL, 4128-CM, R&D Systems).

Techniques: Functional Assay, Generated, Activation Assay, Irradiation, Expressing, Enzyme-linked Immunosorbent Assay, Reporter Assay, Control, In Vitro, Flow Cytometry

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: ATOR-4066 demonstrates strong antitumor activity in syngeneic MC38-expressing human CEACAM5. A, The dose-dependent response of ATOR-4066 was studied in F1 hCD40tgxC57BL/6 mice inoculated with MC38-CEA and treated with the indicated dose of ATOR-4066 or vehicle 6, 10, and 13 days after inoculation, n = 10–27. B, The antitumor effect of ATOR-4066 was compared with an equimolar dose of CD40 mAb and vehicle. C, Flow cytometry analysis and IHC were used to determine CEACAM5 expression in tumors at the indicated time points after cell inoculation in untreated mice, n = 6–10. D, The IHC score was used to determine antibody accumulation in mice treated with 167 μg ATOR-4066, 100 μg of CD40 mAb IHC, or isotype control. Bars represent mean ± SD; each point represents one mouse; n = 5. E and F, Kaplan–Meier plots of mice in which ATOR-4066 treatment (167 µg/dose) was started either 10 days after inoculation ( E ) or 13 days after inoculation ( F ); n = 20. Data represent a representative experiment (from two independent experiments) performed ( B and D ) or pooled data from two ( C , E , F ) independent experiments performed. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. MFI, mean fluorescence intensity.

Article Snippet: In brief, CD40 reporter cells and ATOR-4066 or control bsAb (CD40xCEA bsAb #3) were added to the assay plates before the addition of CEACAM5-transfected CHO cells in the presence or absence of soluble CEACAM5 (0.5, 1, or 5 μg/mL, 4128-CM, R&D Systems).

Techniques: Activity Assay, Expressing, Flow Cytometry, Control, Fluorescence

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: ATOR-4066 alters the transcription profiles of tumors and intratumoral immune cells to a proinflammatory state. A, Transcriptomic alterations induced by ATOR-4066 in blood, tumors, and intratumoral immune cells were studied in F1 hCD40tgxC57BL/6 mice inoculated with MC38-CEACAM5 tumor cells. B and C, PCA illustrated large transcriptomic changes in tumors but not in peripheral blood following ATOR-4066 treatment. D, Gene expression profiles of all DEGs in tumors between the treatment conditions were clustered, and GSEA was performed on each gene cluster. E, GSVA was performed on the top enriched and CD40-related pathways. F, Gene expression of selected canonical inflammatory or immune activation genes. G, The activity of transcription factors (TF) was estimated, and the activity of the top varying transcription factors is shown. H, Gene expression profiles of all DEGs in intratumoral immune cells between the treatment conditions were clustered, and GSEA was performed on each gene cluster. I and J, Immune cell composition was estimated based on CIBERSORTx, and canonical genes for cell populations of immune activation were analyzed. *, P ≤ 0.05; **, P ≤ 0.01. NA, not applicable; Pos., positive; Reg., regulation; VST, variance stabilization transformed.

Article Snippet: In brief, CD40 reporter cells and ATOR-4066 or control bsAb (CD40xCEA bsAb #3) were added to the assay plates before the addition of CEACAM5-transfected CHO cells in the presence or absence of soluble CEACAM5 (0.5, 1, or 5 μg/mL, 4128-CM, R&D Systems).

Techniques: Gene Expression, Activation Assay, Activity Assay, Transformation Assay

Journal: Cancer Immunology Research

Article Title: ATOR-4066, a Bispecific Antibody Targeting CD40 and CEACAM5, Induces Strong Myeloid and T Cell–Dependent Tumor Immunity and Synergizes with PD-1 Blockade

doi: 10.1158/2326-6066.CIR-25-0075

Figure Lengend Snippet: ATOR-4066 in combination with the PD-1 inhibitor enhances the activation of exhausted T cells and controls tumor growth. A and B, ATOR-4066 was evaluated in combination with the anti–PD-1 antibody Opdivo in MLR assays using allogeneic exhausted human primary CD4 + or CD8 + T cells and mature MoDCs. CD4 + or CD8 + T cells with an exhausted phenotype were expanded for 7 days with Dynabeads and then rested overnight in medium. Titrations of either ATOR-4066 and a constant concentration of anti–PD-1 or titrations of anti–PD-1 and a constant concentration of ATOR-4066 in the presence of CEACAM5 beads for cross-linking were used in cultures with a 1:10 mix of mature MoDC cells and exhausted T cells for 7 days before cell culture supernatants were harvested for analyzing IFNγ levels by ELISA. A, Dose–response curves (normalized data, background subtracted, and mean and SD from 9, 14, and 11 donors) are displayed. B, T-cell activation of exhausted CD8 T cells was higher for the combination of ATOR-4066 and PD-1 compared with the sum of the monospecific activity at higher concentrations, exemplified at 1 nmol/L ATOR-4066 and 100 nmol/L anti–PD-1. C and D, To study the in vivo combination treatment effect, F1 hCD40tgxC57BL/6 mice were inoculated with MC38-CEA and treated with 50 μg anti–PD-1 on days 10 and 13 after inoculation and/or 60 μg ATOR-4066 on days 6, 10, and 13 after inoculation. Survival was recorded, and tumor volume was measured. Data represent pooled data from two experiments independently performed. n = 18–20/group. conc., concentration.

Article Snippet: In brief, CD40 reporter cells and ATOR-4066 or control bsAb (CD40xCEA bsAb #3) were added to the assay plates before the addition of CEACAM5-transfected CHO cells in the presence or absence of soluble CEACAM5 (0.5, 1, or 5 μg/mL, 4128-CM, R&D Systems).

Techniques: Activation Assay, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Activity Assay, In Vivo

Journal: Advanced Science

Article Title: A Wearable In‐Pad Diagnostic for the Detection of Disease Biomarkers in Menstruation Blood

doi: 10.1002/advs.202505170

Figure Lengend Snippet: Design of LFAs suitable for biomarker detection in human serum and whole blood in clinically relevant detection windows. a) Design of CEA LFA assay using human CEA‐antibody functionalized 40 nm gold nanoparticles in human serum and b) in fresh, unprocessed human whole blood. c) Design of CA‐125 LFA assay using human CA‐125‐antibody functionalized 40 nm gold nanoparticles in human serum and d) in fresh, unprocessed human whole blood. e) Design of CRP LFA assay using human CRP‐antibody functionalized 20 nm gold nanoparticles in human serum and f) in fresh, unprocessed human whole blood. Assays were performed in triplicate using independent blood donors ( N = 3). LFA images shown in grayscale.

Article Snippet: CRP from human ascites were purchased from Sigma‐Aldrich, and CEA proteins were purchased from Fitzgerald industries international.

Techniques: Biomarker Discovery