尼妥珠单抗原液生产车间工艺设计.docxVIP

尼妥珠单抗原液生产车间设计 摘要 尼妥珠单抗（Nimotuzumab），是一个IgG1 型的人源化单克隆抗体，作用机理为特异性结合EGFR 胞外的结构域的抗原表位 [1]，阻止其活化。临床上主要应用于与放疗联合治疗，以稳固化疗治疗效果。 根据现有尼妥珠单抗氨基酸序列，人工合成尼妥珠单克隆抗体重链与轻链基因，将尼妥珠单抗重链的pSV2-gpt质粒和编码轻链的pSV-hyg质粒转入CHO宿主细胞，获得可稳定表达尼妥珠克隆抗体的工程细胞。 应用细胞灌流培养技术，通过细胞的扩增培养和发酵表达培养，在哺乳动物细胞表达系统利用CHO细胞高效稳定的产生尼妥珠单克隆抗体，然后使用重组protein A层析提纯以及纳米滤膜过滤等纯化工艺进行纯化，再经过无菌灌装单体原液程序产生尼妥珠单抗原液。 针对单克隆抗体生产工艺特性，遵循GMP和国家相关设计规范的要求，根据尼妥珠单克隆抗体的生产工艺流程、设计符合GMP要求的尼妥珠单克隆抗原液生产车间。 关键词：尼妥珠单抗，灌流培养工艺，生产车间设计 Design of Nituzumab Antigen Solution Production Workshop Abstract imotuzumab (Nimotuzumab) is an IgG1 humanized monoclonal antibody that specifically binds to the epitope of the extracellular domain of EGFR [1], preventing its activation. Clinically, it is mainly used in combination with radiotherapy for the treatment of stage III / IV nasopharyngeal carcinoma with positive expression of epidermal growth factor receptor (EGFR). According to the existing amino acid sequence of nimotuzumab, the heavy chain and light chain genes of the monoclonal antibody of nimotuzumab were artificially synthesized, and the pSV2-gpt plasmid of the heavy chain of nimotuzumab and the pSV-hyg plasmid encoding the light chain were transferred into CHO host cells to obtain engineered cells that can stably express Nitozol clone antibodies. Using cell perfusion culture technology, through cell expansion culture and fermentative expression culture, the monoclonal antibody of Nitozol is produced in CHO cells of mammalian cell expression system, which is then purified using recombinant protein A, virus inactivation and nanofiltration membrane filtration. The purification process within the purification, and then aseptically fill the production process of the monoclonal antibody stock solution. According to the characteristics of monoclonal antibody production process, follow the requirements of GMP and relevant national design specifications, according to the production process of Nitrozine monoclonal antibody, design the production workshop of Nitrozine