School of Environmental Sciences, University of Guelph, 50 Stone Road East, Guelph, Ontario N1G 2W1, Canada.
J Agric Food Chem. 2010 Sep 22;58(18):10056-63. doi: 10.1021/jf102284f.
To study the agricultural production of biosimilar antibodies, trastuzumab (Herceptin) was expressed in Nicotiana benthamiana using the magnICON viral-based transient expression system. Immunoblot analyses of crude plant extracts revealed that trastuzumab accumulates within plants mostly in the fully assembled tetrameric form. Purification of trastuzumab from N. benthamiana was achieved using a scheme that combined ammonium sulfate precipitation with affinity chromatography. Following purification, the specificity of the plant-produced trastuzumab for the HER2 receptor was compared with Herceptin and confirmed by western immunoblot. Functional assays revealed that plant-produced trastuzumab and Herceptin have similar in vitro antiproliferative effects on breast cancer cells that overexpress HER2. Results confirm that plants may be developed as an alternative to traditional antibody expression systems for the production of therapeutic mAbs.
为了研究生物类似抗体的农业生产，使用 magnICON 基于病毒的瞬时表达系统在 Nicotiana benthamiana 中表达曲妥珠单抗（赫赛汀）。对粗植物提取物的免疫印迹分析表明，曲妥珠单抗在植物体内主要以完全组装的四聚体形式积累。使用结合硫酸铵沉淀和亲和层析的方案从 N. benthamiana 中纯化曲妥珠单抗。纯化后，通过 Western 免疫印迹比较了植物产生的曲妥珠单抗与赫赛汀对 HER2 受体的特异性，并得到证实。功能测定表明，植物产生的曲妥珠单抗和赫赛汀对过表达 HER2 的乳腺癌细胞具有相似的体外抗增殖作用。结果证实，植物可以作为传统抗体表达系统的替代方法，用于生产治疗性单克隆抗体。
