(10g) A Rapid Screening Platform for Protein Expression to Enable Materials Development
AIChE Annual Meeting
Sunday, November 7, 2021 - 5:18pm to 5:36pm
Herein, a high-throughput E. Coli expression platform has been developed using a low-cost liquid handling robot and open-source software and tested on 17 constructs of interest to demonstrate its wide versatility. Specifically, the genes of interest were inserted into a small library of different DNA plasmids commonly used for biomaterial expression, which include a variety of inducible promoter systems, and further transformed into different cell strains to form a combinatorial expression library that can be tested in well-plate format. Cell growth was monitored by tracking the OD600 value, and a protocol with a simple automated camera was developed and validated such that the platform can be operated without a spectrophotometer. Post-expression, the yield of total protein, via Bradford assay, and of the protein of interest, via dot blot, were quantified to identify promising cell-plasmid combinations that can be further optimized with changes to media, temperature, and time. Reasonable expression conditions can now reliably be found an order of magnitude faster, and conditions for previously un-expressible proteins have been elucidated in several cases. With this large and growing collection of data, it may also be possible to use data-driven methods to predict protein material expression.