Developing media to sustain cell growth and production is an essential and ongoing activity in bioprocess development. Modifications to media can often address host or product-specific challenges, such as low productivity or poor product quality. For other applications, systematic design of new media can facilitate the adoption of new industrially relevant alternative hosts. Despite manifold existing methods, common approaches for optimization often remain time and labor-intensive. We present here a novel approach to conventional media blending that leverages stable, simple, concentrated stock solutions to enable rapid improvement of measurable phenotypes of interest. We applied this modular methodology to generate high-performing media for two phenotypes of interest: biomass accumulation and heterologous protein production, using high-throughput, milliliter-scale batch fermentations of Pichia pastoris as a model system. In addition to these examples, we also created a flexible open-source package for modular blending automation on a low-cost liquid handling system to facilitate wide use of this method. Our modular blending method enables rapid, flexible media development, requiring minimal labor investment and prior knowledge of the host organism, and should enable developing improved media for other hosts and phenotypes of interest.

中文翻译：

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模块化开发能够为替代宿主快速设计媒体

开发培养基以维持细胞生长和生产是生物工艺开发中一项必不可少且持续的活动。对介质的修改通常可以解决宿主或产品特定的挑战，例如低生产率或产品质量差。对于其他应用，新媒体的系统设计可以促进采用新的工业相关替代宿主。尽管存在多种现有方法，但常见的优化方法通常仍然是时间和劳动密集型的。我们在这里提出了一种传统媒体混合的新方法，该方法利用稳定、简单、浓缩的库存解决方案来快速改进可测量的感兴趣的表型。我们应用这种模块化方法为两种感兴趣的表型生成高性能培养基：生物量积累和异源蛋白质生产，使用高通量，毕赤酵母作为模型系统。除了这些示例之外，我们还创建了一个灵活的开源包，用于在低成本液体处理系统上实现模块化混合自动化，以促进该方法的广泛使用。我们的模块化混合方法能够实现快速、灵活的培养基开发，需要最少的劳动力投资和对宿主生物体的先验知识，并且应该能够为其他宿主和感兴趣的表型开发改进的培养基。