Optimization of the hybrid bilayer membrane method for immobilization of avidin on quartz crystal microbalance.

Hybrid bilayer membrane (HBM), comprising a lipid monolayer fused to a hydrophobic self-assembled monolayer (SAM), has a potential capability to provide a convenient tool for the preparation of functionalized sensor surfaces. In this work, the HBM approach was optimized for the preparation of avidin-containing quartz crystal microbalance (QCM) sensor chip which would be available for immobilization of biotinylated molecules. Lipid layer of HBM was composed of background lipid such as egg phosphatidyl choline and biotinylated lipid to which avidin was attached. Highest performance was obtained at 1:1 ratio of the biotinylated lipid and the background lipid, and sensitivity and stability of the resulting sensor chip was comparable to a sensor chip prepared by the conventional carbodiimide reaction. By utilizing the HBM method, construction of a stable avidin sensor chip was achieved within 40 min without any chemical steps. Thus the HBM method was proven to be a convenient and efficient way to immobilize avidin on sensor surfaces.