Hydrogel-stamped membrane protein arrays for high-throughput applications

COLL 159

Sheereen Majd, sheereen@umich.edu1, Ricardo F. Capone, rcapone@umich.edu1, and Michael Mayer, mimayer@umich.edu2. (1) Biomedical Engineering, University of Michigan, room 2149, Lurie Biomedical Engineering Bldg, 1101 beal Ave., ann arbor, MI 48109, (2) Biomedical Engineering and Chemical Engineering, University of Michigan, Lurie Biomedical Engineering Bldg, 1101 beal Ave., ann arbor, MI 48109
Membrane proteins are the targets of almost half of all drugs on the market. Arrays of membrane proteins attract increasing interest to study protein-lipid interactions, protein-membrane protein interactions, and drug-membrane interactions. Fabrication of such arrays is, however, challenging because of the fragile nature of transmembrane proteins. We have recently developed an efficient technique to create multiple copies of membrane arrays using topographically patterned hydrogel stamps. Here, we applied this technique to create membrane microarrays with various embedded transmembrane proteins. Hydrogel stamping makes it possible to form rapidly more than 100 membrane arrays with various compositions while consuming extremely small amounts of material (a few picomoles). The inked posts of a hydrogel stamp store precious biomolecules (in this case lipids, and membrane proteins) in a hydrated and biocompatible environment and deliver them to a substrate in a parallel fashion resulting in an array with spots that may vary both in their lipid and protein compositions. Using FRAP and ligand binding assays, we characterize the resulting membrane protein arrays and examine their usefulness for high-throughput screening assays. The unique properties of the approach presented here may make it a rapid, efficient, and cost-effective strategy for fabrication of membrane protein microarrays for high-throughput applications.