Boonyayothin Wanuttha, Sinnung Sirorut, Shanmugaraj Balamurugan, Abe Yoshito, Strasser Richard, Pavasant Prasit, Phoolcharoen Waranyoo
Research Unit for Plant-Produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand.
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Front Plant Sci. 2021 Jun 23;12:683417. doi: 10.3389/fpls.2021.683417. eCollection 2021.
Denosumab, an anti-receptor activator of nuclear factor-kappa B ligand antibody (anti-RANKL), is a fully human monoclonal antibody (mAb) available for the treatment of osteoporosis. In the present study, an anti-RANKL mAb was transiently expressed using the geminiviral expression system in , and the functional activity of the plant-produced mAb was determined. The highest expression level of the plant-produced mAb was found at 8 days post-infiltration, and it was estimated to be 0.5 mg/g leaf fresh weight. The recombinant mAb from the plant crude extracts was purified by using Protein A affinity column chromatography. The plant-produced mAb demonstrated good affinity binding with human RANKL, as determined by RANKL-ELISA binding. The function of the plant-produced mAb was evaluated . CD14-positive cells isolated from human peripheral blood mononuclear cells (PBMCs) were cultured in the presence of human RANKL and macrophage-colony-stimulating factor (M-CSF) to stimulate osteoclastogenesis. The results demonstrated that plant-produced mAb could significantly decrease the number of osteoclasts compared to commercial denosumab. These results demonstrated that the plant-produced mAb has the potential to inhibit osteoclast differentiation and that it could be considered for osteoporosis treatment.
地诺单抗是一种抗核因子-κB 受体活化因子配体抗体（抗RANKL），是一种可用于治疗骨质疏松症的全人源单克隆抗体（mAb）。在本研究中，利用双生病毒表达系统在[具体植物或生物]中瞬时表达抗RANKL mAb，并测定了植物产生的mAb的功能活性。植物产生的mAb在浸润后8天达到最高表达水平，估计为0.5 mg/g叶鲜重。通过蛋白A亲和柱层析法从植物粗提物中纯化重组mAb。通过RANKL-ELISA结合测定，植物产生的mAb与人RANKL表现出良好的亲和结合。评估了植物产生的mAb的功能。从人外周血单核细胞（PBMCs）中分离出的CD14阳性细胞在人RANKL和巨噬细胞集落刺激因子（M-CSF）存在下培养以刺激破骨细胞生成。结果表明，与市售地诺单抗相比，植物产生的mAb可显著减少破骨细胞数量。这些结果表明，植物产生的mAb具有抑制破骨细胞分化的潜力，可考虑用于骨质疏松症的治疗。
