SummaryThe need for therapeutics to treat a plethora of medical conditions and diseases is on the rise and the demand for alternative approaches to mammalian‐based production systems is increasing. Plant‐based strategies provide a safe and effective alternative to produce biological drugs but have yet to enter mainstream manufacturing at a competitive level. Limitations associated with batch consistency and target protein production levels are present; however, strategies to overcome these challenges are underway. In this study, we apply state‐of‐the‐art mass spectrometry‐based proteomics to define proteome remodelling of the plant following agroinfiltration with bacteria grown under shake flask or bioreactor conditions. We observed distinct signatures of bacterial protein production corresponding to the different growth conditions that directly influence the plant defence responses and target protein production on a temporal axis. Our integration of proteomic profiling with small molecule detection and quantification reveals the fluctuation of secondary metabolite production over time to provide new insight into the complexities of dual system modulation in molecular pharming. Our findings suggest that bioreactor bacterial growth may promote evasion of early plant defence responses towards Agrobacterium tumefaciens (updated nomenclature to Rhizobium radiobacter). Furthermore, we uncover and explore specific targets for genetic manipulation to suppress host defences and increase recombinant protein production in molecular pharming.

中文翻译：

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本塞姆氏烟草的细菌生长介导的系统重塑定义了分子制药中靶蛋白生产的独特特征

摘要对治疗多种医疗状况和疾病的治疗方法的需求不断增加，并且对基于哺乳动物的生产系统的替代方法的需求也在增加。基于植物的策略为生产生物药物提供了安全有效的替代方案，但尚未以竞争水平进入主流制造。存在与批次一致性和目标蛋白质生产水平相关的限制；然而，克服这些挑战的战略正在进行中。在这项研究中，我们应用最先进的基于质谱的蛋白质组学来定义在摇瓶或生物反应器条件下生长的细菌进行农杆菌渗透后植物的蛋白质组重塑。我们观察到与不同生长条件相对应的细菌蛋白质生产的不同特征，这些特征直接影响时间轴上的植物防御反应和目标蛋白质生产。我们将蛋白质组分析与小分子检测和定量相结合，揭示了次级代谢物产生随时间的波动，为分子制药中双系统调节的复杂性提供了新的见解。我们的研究结果表明，生物反应器细菌的生长可能会促进植物逃避早期防御反应根癌农杆菌（将命名法更新为放射根瘤菌）。此外，我们发现并探索基因操作的特定靶标，以抑制宿主防御并增加分子制药中重组蛋白的产量。