Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Methods Mol Biol. 2022;2411:37-62. doi: 10.1007/978-1-0716-1888-2_2.
For more than three decades, mammalian cells have been the host par excellence for the recombinant protein production for therapeutic purposes in humans. Due to the high cost of media and other supplies used for cell growth, initially this expression platform was only used for the production of proteins of pharmaceutical importance including antibodies. However, large biotechnological companies that used this platform continued research to improve its technical and economic feasibility. The main qualitative improvement was obtained when individual cells could be cultured in a liquid medium similar to bacteria and yeast cultures. Another important innovation for growing cells in suspension was the improvement in chemically defined media that does not contain macromolecules; they were cheaper to culture as any other microbial media. These scientific milestones have reduced the cost of mammalian cell culture and their use in obtaining proteins for veterinary use. The ease of working with mammalian cell culture has permitted the use of this expression platform to produce active pharmaceutic ingredients for veterinary vaccines. In this chapter, the protocol to obtain recombinant mammalian cell lines will be described.
三十多年来,哺乳动物细胞一直是人类治疗性重组蛋白生产的首选宿主。由于用于细胞生长的培养基和其他耗材成本高昂,最初该表达平台仅用于生产具有药物重要性的蛋白质,包括抗体。然而,使用该平台的大型生物技术公司继续进行研究,以提高其技术和经济可行性。当可以将单个细胞在类似于细菌和酵母培养物的液体培养基中培养时,获得了主要的定性改进。另一个用于悬浮细胞生长的重要创新是在化学成分确定的培养基方面的改进,该培养基不含大分子物质;与其他微生物培养基相比,它们的培养成本更低。这些科学里程碑降低了哺乳动物细胞培养的成本,并使其可用于获得兽用蛋白。由于易于操作哺乳动物细胞培养,因此可以使用该表达平台生产用于兽用疫苗的活性药物成分。在本章中,将描述获得重组哺乳动物细胞系的方案。