High‐throughput evolutionary optimization of the induction medium towards recombinant protein production in BY‐2 tobacco,Biotechnology and Bioengineering

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High‐throughput evolutionary optimization of the induction medium towards recombinant protein production in BY‐2 tobacco
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2020-10-10 , DOI: 10.1002/bit.27594
Jan Sadoch ₁ , Monika Pyc ₁ , Anna Urbanowicz ₂ , Adam Iglewski ₃ , Radosław Pilarski ₁

Affiliation

Bright yellow (BY‐2) tobacco cells combined with the XVE chemically inducible system are one of the most promising plant‐based platforms for recombinant protein production. This offers a range of benefits, including the separation of the cell growth and heterologous gene expression, lack of risk of infecting the end product with prions and human viruses or appropriate protein glycosylation and folding. However, low protein productivity remains a major obstacle that limits the extensive commercialization of bioproduction in plants. A number of molecular, cell culture and down processing approaches have been made to overcome this problem. Media development for the specific nutritional and hormonal requirements of transgenic plant cells is one of the most efficient cell‐culture approaches. We optimized the induction medium towards recombinant protein production in BY‐2 and demonstrated the usefulness of evolutionary medium optimization for high‐yield protein production in liquid plant cultures. A reliable XVE/GFP model, parallel conducting experiments in a microscale on 96‐well plates, and dedicated Gene Game evolutionary optimization software allowed for an effective search of 76¹¹ possible solutions of 11‐component media. Within the 4608 formulations tested, the Induct X medium was found with a significant 107.14% increase in protein expression in relation to the standard BY‐2 medium.

中文翻译：

BY-2 烟草中重组蛋白生产诱导培养基的高通量进化优化

亮黄色 (BY-2) 烟草细胞与 XVE 化学诱导系统相结合是最有前途的植物重组蛋白生产平台之一。这提供了一系列好处，包括细胞生长和异源基因表达的分离、最终产品没有感染朊病毒和人类病毒的风险或适当的蛋白质糖基化和折叠。然而，低蛋白质生产率仍然是限制植物生物生产广泛商业化的主要障碍。已经采取了许多分子、细胞培养和羽绒加工方法来克服这个问题。针对转基因植物细胞特定营养和激素需求的培养基开发是最有效的细胞培养方法之一。我们优化了 BY-2 中重组蛋白生产的诱导培养基，并证明了进化培养基优化对于液体植物培养物中高产蛋白生产的有用性。可靠的 XVE/GFP 模型、在 96 孔板上进行微尺度平行实验以及专用的 Gene Game 进化优化软件，可以有效搜索 11 组分培养基的 76 ^{11 种}可能的解决方案。在测试的 4608 种配方中，发现 Induct X 培养基的蛋白质表达相对于标准 BY-2 培养基显着增加了 107.14%。

更新日期：2020-10-10

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