Guiding In Vivo Protein Expression with eProtein Discovery™ CFPS for Disulfide-Rich Proteins

The eProtein Discovery System simplifies the transition from cell-free protein synthesis (CFPS) to in vivo. E. coli expression, especially for intricate proteins with disulfide bonds. By evaluating numerous protein variants and up to seven solubility tags, researchers can efficiently identify optimal construct designs.

By reading this application note you’ll gain insight into the following:

• A recent study where 15 proteins were examined. PDI (protein disulfide isomerase) and GSSG (oxidized glutathione) proved crucial for disulfide bond formation and maximizing yields.
• Screening results strongly correlated with CFPS yields, validating the system's reliability.
• SHuffle® T7 Express strain demonstrated the highest correlation with CFPS results, likely due to its inherent disulfide bond formation capabilities, mimicking the effects of PDI/GSSG.
• In contrast, BL21(DE3) strain and the absence of these additives yielded lower protein levels.

This application note underscores the value of CFPS as a powerful tool for optimizing protein expression and guiding in vivo scale-up, reducing the need for multiple iterations in plasmid design and strain selection. The eProtein Discovery System ultimately accelerates protein production, providing researchers with a dependable solution for obtaining functional proteins.