Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Department of Otorhinolaryngology-Head and Neck Surgery, Chung-Ang University College of Medicine, Seoul 156-756, Korea.
Int J Mol Sci. 2020 Feb 17;21(4):1354. doi: 10.3390/ijms21041354.
Overexpression of human epidermal growth factor receptor type 2 (HER2) is considered as a prognostic factor of breast cancer, which is positively associated with recurrence when cancer metastasizes to the lymph nodes. Here, we expressed the single variable domain on a heavy chain (VHH) form of anti-HER2 camelid single domain antibody in tobacco plants and compared its in vitro anticancer activities with the anti-HER2 full size antibody. The gene expression cassette containing anti-HER2 camelid single domain antibody VHH fused to human IgG Fc region with KDEL endoplasmic reticulum (ER) (VHH-FcK) was transferred into the tobacco plant via the -mediated transformation. The transformants were screened with polymerase chain reaction and Western blot analyses. Enzyme-linked immunosorbent assay (ELISA) confirmed the binding of the purified anti-HER2 VHH-FcK to the HER2-positive breast cancer cell line, SK-BR-3. Migration assay results confirmed anticancer activity of the plant-derived anticancer camelid single chain antibody. Taken together, we confirmed the possibility of using anti-HER2 VHH-FcK as a therapeutic anticancer agent, which can be expressed and assembled and purified from a plant expression system as an alternative antibody production system.
人表皮生长因子受体 2（HER2）的过表达被认为是乳腺癌的预后因素，当癌症转移到淋巴结时，它与复发呈正相关。在这里，我们在烟草植物中表达了抗 HER2 骆驼单域抗体的重链（VHH）形式的单可变结构域，并将其体外抗癌活性与抗 HER2 全尺寸抗体进行了比较。含有抗 HER2 骆驼单域抗体 VHH 与人 IgG Fc 区融合的 KDEL 内质网（ER）（VHH-FcK）的基因表达盒通过介导的转化转入烟草植物。通过聚合酶链反应和 Western blot 分析筛选转化体。酶联免疫吸附测定（ELISA）证实了纯化的抗 HER2 VHH-FcK 与 HER2 阳性乳腺癌细胞系 SK-BR-3 的结合。迁移试验结果证实了植物来源的抗癌骆驼单链抗体的抗癌活性。综上所述，我们证实了使用抗 HER2 VHH-FcK 作为治疗性抗癌药物的可能性，它可以从植物表达系统中表达、组装和纯化，作为替代抗体生产系统。
