肿瘤免疫治疗中抗tigit和CD155单克隆抗体的制备

Y. Duan, Yan-lin Bian, Jianjia Zhu
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引用次数: 0

摘要

许多研究证实,人类脊髓灰质炎病毒受体(PVR;CD155)与肿瘤细胞迁移、侵袭和肿瘤进展有关。PVR受体结合其配体T细胞Ig和ITIM结构域(TIGIT)抑制T和NK细胞的功能,从而使肿瘤逃避免疫监视。本研究通过哺乳动物瞬时转染系统表达抗cd155和抗tigit两种IgG1单克隆抗体,然后在体外评估抗体依赖细胞介导的细胞毒性、抗体结合亲和力和抗肿瘤功效。本研究采用蛋白A亲和层析法纯化抗体。分析方法包括免疫印迹法、酶联免疫吸附法和流式细胞术。我们的数据表明,这两种单克隆抗体的纯度均高于90%,并且与抗原结合紧密,解离常数(K d)和50%有效浓度(EC50)低于微摩尔范围。最值得注意的是,这些抗体在体外促进免疫细胞的抗肿瘤活性。因此,本研究为后续体内实验进一步评价奠定了基础。
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Generating Anti-TIGIT and CD155 Monoclonal Antibodies for Tumor Immunotherapy
Many studies have confirmed that the human poliovirus receptor (PVR; CD155) is related to tumor cell migration, invasion, and thus tumor progression. A PVR receptor binds its ligand T cell Ig and the ITIM domain (TIGIT) to inhibit the function of T and NK cells, thereby allowing tumors to evade immune surveillance. In this study, two IgG1 monoclonal antibodies, anti-CD155 and anti-TIGIT, were expressed by the mammalian transient transfection system, then, antibody-dependent cell-mediated cytotoxicity, antibody-binding affinity, and antitumor efficacy were evaluated subsequently in vitro. In this work, protein A affinity chromatography was used for antibodies' purification. Analysis methods included Western blot, enzyme-linked immunosorbent assay, and flow cytometry. Our data suggested that both the two monoclonal antibodies have a purity of higher than 90%, and bound tightly to the antigen with dissociation constant (K d) and 50% effective concentrations (EC50) below micromolar range. Most notably, these antibodies promote antitumor activity of immune cells in vitro. Therefore, our study laid down the foundation for subsequent in vivo experiments for further evaluation.
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审稿时长
15 weeks
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