Figure S4 from A Novel Designed Anti–PD-L1/OX40 Bispecific Antibody Augments Both Peripheral and Tumor-Associated Immune Responses for Boosting Antitumor Immunity

作者：Baocun Li, Shiyong Gong, Nianying Zhang, Beilei Shi, Zhou Lv, Yu Zhang, Naren Gaowa, Liqin Dong, Danqing Wu, Jianfu Wu, Fan Liu, Rui Zhang, Ramin Behzadigohar, Vinod Ganju, Chengbin Wu, Xuan Wu · 年份：2026 · DOI：10.1158/1535-7163.32715407 · 研究领域：Monoclonal and Polyclonal Antibodies Research、CAR-T cell therapy research、Cancer Immunotherapy and Biomarkers

<p>Supplementary Fig.4. Characterization of EMB-09 binding. a Antibodies binding to CHO cells overexpressing OX40 (left), CHO-PD-L1 cells (right). b Antibodies binding to resting CD4+ or activated CD4+ T cells by flow cytometry. PD-L1 and OX40 expression on activated CD4+ T cells stimulated with anti-CD3/CD28 for 48h. c The binding kinetic constants of PD-L1/OX40 to soluble PD-L1 antigens (left) or to soluble OX40 (right) was determined by BLI. d Concomitant binding of PD-L1 and OX40 by EMB-09. Plates were coated with recombinant hOX40 protein (left) or hPD-L1 protein (right), antibody binding to each coated antigen were detected with either biotinylated PD-L1 or OX40 respectively. 44-16 is the parental anti-OX40 mAb of EMB-09, 86-64 is the parental anti-PD-L1 mAb of EMB-09. e The blocking properties of EMB-09 to PD-L1/PD1 interaction were measured by protein based receptor blocking assay and cell based PD-L1/PD1 reporter assay. Left: The blocking of PD1 protein with mFc tag to PD-L1 expressing CHO-K1 in the presence of various concentrations of EMB-09, parental anti-PD-L1 mAb (86-64), or isotype control (hIgG); Right: The PD-L1 blocking efficacy of EMB-09 or the parental anti-PD-L1 mAb was assessed by measuring downstream NFAT luciferase activity in a co-culture system consisting of CHO-PD-L1 and Jurkat-PD-1-NFAT-Luc.</p>