(672c) Nanoscale Surface Patterning of Multiple Proteins Using Photoactivation
Patterned protein surfaces are important for studies of cellular interactions with the extracellular matrix. However, next-generation patterned protein surfaces require improvements to decrease feature size, place different proteins in registry, and control orientation of the active domains to maintain full activity. Here, we describe a photochemical method that uses photoactivated ligands for covalent immobilization of fusion proteins and we demonstrate the patterning of two different proteins with sub-micron feature sizes in registry. We designed a self-assembled monolayer that presented photoprotected SnapTag ligands, and patterned UV illumination selectively activated covalent immobilization of SnapTag fusion proteins. We patterned a fluorescent protein with feature sizes of 400 nm over a 1 mm2 area, and sequential patterning placed nanoscale patterns of different proteins in close proximity. We applied this technique to pattern multiple cell ligands on the nano and microscale, and observed the response of human cell lines to different ligand patterns. This method enables the preparation of surfaces having multiple proteins independently patterned with control over the densities and activities of proteins and will be important in studies of cell signaling from the extracellular matrix.