A beginner’s guide to design of experiments for life sciences using Spreadsheet DoE

Life science research frequently involves optimizing experimental conditions to maximize product yield, ensuring the research pipeline can progress efficiently. For instance, in biochemistry, many processes in the protein production and characterization pipeline need optimizing such as cloning, protein expression, protein purification, and a range of structural biology techniques and functional assays. Design of Experiments (DoE) is an approach that designs a set of experiments that varies all factors at the same time for rapid process optimization, however, this approach presents a challenge due to statistical complexities and high software costs. To address this, DoE is introduced with Spreadsheet DoE, an open access user-friendly educational factorial design tool that provides step-by-step guidance and transparent spreadsheet calculations. Protein expression optimization in Escherichia coli is used as an example, and other biochemical optimizations are also explored. This guide along with Spreadsheet DoE aims to encourage researchers with minimal statistical background to effectively employ DoE methodologies in their life science studies.