Zhang Richard Yi, Shen Wenyan David
Accurus Biosciences Inc, Dublin, CA, USA.
Methods Mol Biol. 2012;907:341-58. doi: 10.1007/978-1-61779-974-7_20.
In the past two decades, the production levels for monoclonal antibodies in mammalian expression systems have improved dramatically. Single cell productivity for monoclonal antibodies has increased 20-50 fold due to the improvements in expression hosts, expression vectors, cell culture media, and production processes. However, most of these improvements are proprietary to large pharmaceutical/biotech companies and involve large steel-tank bioreactors. Therefore, these processes are difficult for small companies and academic labs to reproduce. Transient expression in mammalian cells has recently been used very widely for monoclonal antibody expression. Cell line and expression vector engineering increased expression levels to several hundred milligrams per liter. The availability of highly effective transfection reagents and disposable bioreactors make the transient expression process an efficient and cost-effective way to make recombinant antibodies in large quantity. Here, we describe the protocols for small- to mid-scale transient expression of monoclonal antibodies in shake-flasks and for large-scale production in WAVE bioreactors.
在过去二十年中，哺乳动物表达系统中单克隆抗体的生产水平有了显著提高。由于表达宿主、表达载体、细胞培养基和生产工艺的改进，单克隆抗体的单细胞生产力提高了20至50倍。然而，这些改进大多为大型制药/生物技术公司所专有，且涉及大型钢罐生物反应器。因此，小公司和学术实验室很难重现这些工艺。哺乳动物细胞中的瞬时表达最近已被广泛用于单克隆抗体的表达。细胞系和表达载体工程使表达水平提高到每升数百毫克。高效转染试剂和一次性生物反应器的可用性使瞬时表达过程成为大量生产重组抗体的一种高效且经济高效的方法。在此，我们描述了在摇瓶中进行中小规模单克隆抗体瞬时表达以及在WAVE生物反应器中进行大规模生产的方案。
