Kadwell Sue H, Overton Laurie K
Department of Biological Sciences, Molecular Discovery Research, Platform Technology and Science, GlaxoSmithKline, Research Triangle Park, NC, USA.
Methods Mol Biol. 2016;1350:263-84. doi: 10.1007/978-1-4939-3043-2_12.
Many types of disposable bioreactors for protein expression in insect and mammalian cells are now available. They differ in design, capacity, and sensor options, with many selections available for either rocking platform, orbitally shaken, pneumatically mixed, or stirred-tank bioreactors lined with an integral disposable bag (Shukla and Gottschalk, Trends Biotechnol 31(3):147-154, 2013). WAVE Bioreactors™ were among the first disposable systems to be developed (Singh, Cytotechnology 30:149-158, 1999). Since their commercialization in 1999, Wave Bioreactors have become routinely used in many laboratories due to their ease of operation, limited utility requirements, and protein expression levels comparability to traditional stirred-tank bioreactors. Wave Bioreactors are designed to use a presterilized Cellbag™, which is attached to a rocking platform and inflated with filtered air provided by the bioreactor unit. The Cellbag can be filled with medium and cells and maintained at a set temperature. The rocking motion, which is adjusted through angle and rock speed settings, provides mixing of oxygen (and CO2, which is used to control pH in mammalian cell cultures) from the headspace created in the inflated Cellbag with the cell culture medium and cells. This rocking motion can be adjusted to prevent cell shear damage. Dissolved oxygen and pH can be monitored during scale-up, and samples can be easily removed to monitor other parameters. Insect and mammalian cells grow very well in Wave Bioreactors (Shukla and Gottschalk, Trends Biotechnol 31(3):147-154, 2013). Combining Wave Bioreactor cell growth capabilities with recombinant baculoviruses engineered for insect or mammalian cell expression has proven to be a powerful tool for rapid production of a wide range of proteins.
现在有多种用于昆虫和哺乳动物细胞中蛋白质表达的一次性生物反应器。它们在设计、容量和传感器选项上有所不同，对于摇床平台、轨道振荡、气动混合或内衬一体式一次性袋子的搅拌罐生物反应器都有许多选择（舒克拉和戈特沙尔克，《生物技术趋势》31(3):147 - 154，2013年）。WAVE生物反应器™是最早开发的一次性系统之一（辛格，《细胞技术》30:149 - 158，1999年）。自1999年商业化以来，由于其操作简便、实用要求有限且蛋白质表达水平与传统搅拌罐生物反应器相当，Wave生物反应器已在许多实验室中常规使用。Wave生物反应器设计用于使用预先灭菌的细胞袋™，该细胞袋连接到摇床平台，并由生物反应器单元提供的过滤空气充气。细胞袋可以填充培养基和细胞，并保持在设定温度。通过角度和摇动速度设置进行调节的摇动运动，使来自充气细胞袋中产生的顶空的氧气（以及用于控制哺乳动物细胞培养物中pH值的二氧化碳）与细胞培养基和细胞混合。这种摇动运动可以进行调节以防止细胞剪切损伤。在放大过程中可以监测溶解氧和pH值，并且可以轻松取出样品以监测其他参数。昆虫和哺乳动物细胞在Wave生物反应器中生长良好（舒克拉和戈特沙尔克，《生物技术趋势》31(3):147 - 154，2013年）。事实证明，将Wave生物反应器的细胞生长能力与为昆虫或哺乳动物细胞表达而设计的重组杆状病毒相结合，是快速生产多种蛋白质的强大工具。
