Departments of Pathology and Medicine, College of Medicine, Chung-Ang University, Seoul 06974, Korea.
Departments of Medicine, College of Medicine, Chung-Ang University, Seoul 06974, Korea.
BMB Rep. 2020 Apr;53(4):229-233. doi: 10.5483/BMBRep.2020.53.4.106.
The anti-colorectal cancer monoclonal antibody CO17-1A (mAb CO), which recognizes the tumor-associated antigen EpCAM, was expressed in transgenic Arabidopsis plants. PCR and western blot analyses showed the insertion and expression of heavy chain (HC)/HC fused to the KDEL ER retention modif (HCK) and light chain (LC) of mAb CO and mAb CO with HCK (mAb COK) in Arabidopsis transformants. Both plantderived mAbP CO and mAbP COK were purified from a biomass of approximately 1,000 seedlings grown in a greenhouse. In sandwich ELISA, both mAbP CO showed a slightly higher binding affinity for the target, EpCAM, compared to mAbM CO. In cell ELISA, both mAbsP COs showed binding affinity to the human colorectal cancer cell line SW480. Furthermore, mAbM CO, mAbP CO, and mAbP COK exhibited dose and timedependent regression effects on SW480 cells in vitro. In summation, both mAbP CO and mAbP COK, expressed in Arabidopsis, recognized the target antigen EpCAM and showed anti-proliferative activity against human colorectal cancer cells. [BMB Reports 2020; 53(4): 229-233].
抗结直肠癌单克隆抗体 CO17-1A(mAb CO)识别肿瘤相关抗原 EpCAM,在转基因拟南芥植物中表达。PCR 和 Western blot 分析表明,mAb CO 和 mAb CO 与 HCK(mAb COK)的重链(HC)/HC 融合到 KDEL ER 保留修饰(HCK)和轻链(LC)在拟南芥转化体中插入和表达。两种植物衍生的 mAbP CO 和 mAbP COK 均从温室中生长的约 1000 株幼苗的生物质中纯化而来。在夹心 ELISA 中,与 mAbM CO 相比,两种 mAbP CO 对靶标 EpCAM 的结合亲和力略高。在细胞 ELISA 中,两种 mAbsP CO 均与人类结直肠癌细胞系 SW480 结合。此外,mAbM CO、mAbP CO 和 mAbP COK 在体外对 SW480 细胞表现出剂量和时间依赖性的回归作用。总之,在拟南芥中表达的两种 mAbP CO 和 mAbP COK 均识别靶抗原 EpCAM,并对人结直肠癌细胞表现出抗增殖活性。[BMB 报告 2020;53(4):229-233]。