Synthetic Protein Switches: Theoretical and Experimental Considerations

Abstract

Synthetic protein switches with tailored response functions are finding increasing applications as tools in basic research and biotechnology. With a number of successful design strategies emerging, the construction of synthetic protein switches still frequently necessitates an integrated approach that combines detailed biochemical and biophysical characterization in combination with high-throughput screening to construct tailored synthetic protein switches. This is increasingly complemented by computational strategies that aim to reduce the need for costly empirical optimization and thus facilitate the protein design process. Successful computational design approaches range from analyzing phylogenetic data to infer useful structural, biophysical, and biochemical information to modeling the structure and function of proteins ab initio. The following chapter provides an overview over the theoretical considerations and experimental approaches that have been successful applied in the construction of synthetic protein switches.

Engineering Protein Switches for Rapid Diagnostic Tests

“…Inspiration can be taken from nature, where signaling is underpinned by protein switches that sense and respond to molecular inputs . Engineered protein switches have been designed for synthetic biology and sensing applications and could be transformational as a rapid alternative to laboratory immunoassays. − Immunoassays typically utilize separate reagents for analyte recognition and signal generation, necessitating multiple immobilization and wash steps that prohibit direct point-of-need use (Figure A) . Microfluidic lab-on-a-chip systems can automate the steps, but few, like the Abbott iSTAT point-of-care analyzer, have been commercially successful .…”