DNA-tagged protein libraries for high-throughput biophysical data generation

Models of protein function are limited by the availability of quantitative measurements reported in physical units under uniform conditions. Coupling biophysical assays to sequencing provides a proven route to generating such data at scale, but requires each protein in a library to be associated with a recoverable DNA identifier. Existing approaches to directly couple protein molecules to a DNA tag in vitro are limited by yield, the proteins they accommodate, and protocol complexity. We present a method in development for generating DNA-tagged protein libraries by compartmentalized DNA amplification, expression, and covalent protein-DNA conjugation. The protocol is completed in one day and imposes no size restriction beyond in vitro expression. Our approach could broaden the range of proteins accessible to existing high-throughput assays, such as quantification of folding free energies, and enable new methods, such as library-on-library measurement of protein-protein interactions.