Abstract：
Mammalian cell lines are one of the best options when it comes to the production of complex proteins requiring specific glycosylation patterns. Plasmid DNA transfection and stable cell lines are frequently used for recombinant protein production, but they are expensive at large scale or can become time-consuming, respectively. The BacMam baculovirus (BV) is a safe and cost-effective platform to produce recombinant proteins in mammalian cells. The process of generating BacMam BVs is straightforward and similar to the generation of \"insect\" BVs, with different commercially available platforms. Although there are several protocols that describe recombinant protein expression with the BacMam BV in adherent cell lines, limited information is available on suspension cells. Therefore, it is of relevance to define the conditions to produce recombinant proteins in suspension cell cultures with BacMam BVs that facilitate bioprocess transfer to larger volumes. Here, we describe a method to generate a high titer BacMam BV stock and produce recombinant proteins in suspension HEK293 cells.
摘要：
哺乳动物细胞系是生产需要特定糖基化模式的复杂蛋白质的最佳选择之一。质粒DNA转染和稳定细胞系经常用于重组蛋白生产，但是它们在大规模上很昂贵，或者可能变得耗时，分别。BacMam杆状病毒（BV）是在哺乳动物细胞中生产重组蛋白的安全且具有成本效益的平台。生成BacMamBV的过程很简单，类似于“昆虫”BV的生成，不同的商业平台。尽管有几种方案描述了在贴壁细胞系中BacMamBV的重组蛋白表达，关于悬浮细胞的信息有限。因此,定义在具有BacMamBV的悬浮细胞培养物中产生重组蛋白的条件是相关的,其促进生物过程转移到更大体积。这里,我们描述了在悬浮HEK293细胞中产生高滴度BacMamBV原液并产生重组蛋白的方法。
