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: Directly after thawing, DTCs were counted using a NucleoCounter NC-200 (ChemoMetec), stained for CEACAM5 and CD40, and analyzed by flow cytometry to confirm target expression at baseline.

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