Abstract
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.