Facile Protein Immobilization Using Engineered Surface-Active Biofilm Proteins

Immobilization of enzymes and other biomolecules to surfaces is critically important for biotechnology, with important applications in sensing and controlled delivery of molecular species for analytical or biomedical purposes. The presentation of protein recognition elements in a way that avoids denaturation and nonspecific interactions while maintaining the accessibility of the active site is a challenge for which no general solution has been found. Here we present a robust, facile method for immobilization of any protein to a surface using engineered protein building blocks. By functionalizing an interfacial protein, BslA, with peptides (SpyTag and SnoopTag) that spontaneously react with their cognate protein partners (SpyCatcher and SnoopCatcher), we are able to create patterned surfaces of protein monolayers displaying reactive tags. We demonstrate that these surfaces can be functionalized rapidly, spontaneously, and specifically with proteins of interest attached to SpyCatcher or SnoopCatcher. This m...