PT  - JOURNAL ARTICLE
AU  - Romel Menacho-Melgar
AU  - Jennifer N. Hennigan
AU  - Michael D. Lynch
TI  - Optimization of Phosphate Limited Autoinduction Broth for 2-Stage Heterologous Protein Expression in <em>E. coli</em>
AID  - 10.1101/2021.06.08.447502
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.06.08.447502
4099  - http://biorxiv.org/content/early/2021/06/08/2021.06.08.447502.short
4100  - http://biorxiv.org/content/early/2021/06/08/2021.06.08.447502.full
AB  - Autoinducible, 2-stage protein expression leveraging phosphate inducible promoters has been recently shown to enable not only high protein titers but also consistent performance across scales from screening systems (microtiter plates) to instrumented bioreactors. However, to date small scale production using microtiter plates and shake flasks rely on a complex autoinduction broth (AB) that requires making numerous media components, not all amenable to autoclaving. In this report, we develop a simpler media formulation (AB-2) with just a few autoclavable components. We show that AB-2 is robust to small changes in its composition and performs equally, if not better, than AB across different scales. AB-2 will facilitate adoption of phosphate limited 2-stage protein expression protocols.Competing Interest StatementM.D. Lynch has a financial interest in DMC Biotechnologies, Inc. M.D. Lynch, J.N. Hennigan and R. Menacho-Melgar have a financial interest in Roke Biotechnologies, Inc.