Bulaon Christine Joy I, Khorattanakulchai Narach, Rattanapisit Kaewta, Sun Hongyan, Pisuttinusart Nuttapat, Strasser Richard, Tanaka Shiho, Soon-Shiong Patrick, Phoolcharoen Waranyoo
Center of Excellence in Plant-Produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand.
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Front Plant Sci. 2023 Aug 29;14:1149455. doi: 10.3389/fpls.2023.1149455. eCollection 2023.
Cytotoxic T lymphocyte-associated protein 4 (CTLA-4) is an immune checkpoint regulator exclusively expressed on T cells that obstructs the cell's effector functions. Ipilimumab (Yervoy®), a CTLA-4 blocking antibody, emerged as a notable breakthrough in modern cancer treatment, showing upfront clinical benefits in multiple carcinomas. However, the exhilarating cost of checkpoint blockade therapy is discouraging and even utmost prominent in developing countries. Thereby, affordability of cancer care has become a point of emphasis in drug development pipelines. Plant expression system blossomed as a cutting-edge platform for rapid, facile to scale-up, and economical production of recombinant therapeutics. Here, we describe the production of an anti-CTLA-4 2C8 antibody in . ELISA and bio-layer interferometry were used to analyze antigen binding and binding kinetics. Anticancer responses in vivo were evaluated using knocked-in mice implanted with syngeneic colon tumor. At 4 days post-infiltration, the antibody was transiently expressed in plants with yields of up to 39.65 ± 8.42 μg/g fresh weight. Plant-produced 2C8 binds to both human and murine CTLA-4, and the plant-produced IgG1 also binds to human FcγRIIIa (V158). In addition, the plant-produced 2C8 monoclonal antibody is as effective as Yervoy® in inhibiting tumor growth in vivo. In conclusion, our study underlines the applicability of plant platform to produce functional therapeutic antibodies with promising potential in cancer immunotherapy.
细胞毒性T淋巴细胞相关蛋白4（CTLA-4）是一种仅在T细胞上表达的免疫检查点调节因子，它会阻碍细胞的效应功能。伊匹单抗（Yervoy®）是一种CTLA-4阻断抗体，成为现代癌症治疗中的一项显著突破，在多种癌症中显示出初步的临床益处。然而，检查点阻断疗法高昂的成本令人却步，在发展中国家尤为突出。因此，癌症治疗的可负担性已成为药物研发流程中的一个重点。植物表达系统作为一个前沿平台蓬勃发展，可用于快速、易于扩大规模且经济地生产重组治疗药物。在此，我们描述了在植物中生产抗CTLA-4 2C8抗体的过程。采用酶联免疫吸附测定（ELISA）和生物层干涉术分析抗原结合及结合动力学。使用植入同基因结肠肿瘤的基因敲入小鼠评估体内抗癌反应。浸润后4天，该抗体在植物中瞬时表达，产量高达39.65±8.42μg/g鲜重。植物生产的2C8能与人及小鼠的CTLA-4结合，且植物生产的IgG1也能与人FcγRIIIa（V158）结合。此外，植物生产的2C8单克隆抗体在体内抑制肿瘤生长方面与Yervoy®一样有效。总之，我们的研究强调了植物平台在生产具有癌症免疫治疗潜力的功能性治疗性抗体方面的适用性。
