We report a new methodology to pattern and immobilize ligands quantitatively to planarsupported fluid lipid bilayers. We pattern bilayer surfaces by combining a new microfluidic lithography strategy and immobilize ligands to the bilyaer by use of an electroactive and chemoselective interfacial reaction strategy. The bilayers were characterized by electrochemistry, fluorescence microscopy and determined to be fluid, an important property in mimicking the natural cell membrane, by fluorescence recovery after photobleaching (FRAP). Electroactive vesicles were first generated by the incorporation of hydroquinone-tethered alkane (H2Q) with egg palmitoyl-oleoyl phosphatidylcholine (egg-POPC) and then subsequently fused to form a patterned fluid bilayer on a conducting gold self assembled monolayer surface. The H2Q can be electrochemically oxidized to the corresponding quinone (Q), followed by conjugation with an oxyamine-functionalized (RONH2) ligand to the bilayer. This strategy provides a synergistic method to pattern and present a number of ligands or biomolecules from the bilayer surface for evaluation of enzymatic or protein binding to biomembranes. The oxime-product is also electroactive and can be monitored by electrochemistry to precisely determine the amount of ligand immobilization. As examples, an oxyamine-tethered glucose was synthesized and immobilized to patterned bilayers followed by TRITC-conjugated Concanavalin A (ConA), generating a ligand array visualized by fluorescence microscopy.

Selective tethering of ligands and proteins to a microfluidic patterned electroactive fluid lipid bilayer array

240th National Meeting (2010)

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