Reproducing bench-scale cell growth and productivity

Reproducing, exchanging, comparing, and building on each other’s work is foundational to technology advances.1 Advancing biotechnology calls for reliable reuse of engineered strains.2 Reliable reuse of engineered strains requires reproducible growth and productivity. To demonstrate reproducibility for biotechnology, we identified the experimental factors that have the greatest effect on the growth and productivity of our engineered strains.3–6 We present a draft of a Minimum Information Standard for Engineered Organism Experiments (MIEO) based on this method. We evaluated the effect of 22 factors on Escherichia coli (E. coli) engineered to produce the small molecule lycopene, and 18 factors on E. coli engineered to produce red fluorescent protein (RFP). Container geometry and shaking had the greatest effect on product titer and yield. We reproduced our results under two different conditions of reproducibility:7 conditions of use (different fractional factorial experiments), and time (48 biological replicates performed on 12 different days over four months).