Immobilization of Olfactory Receptors Carried by Nanosomes onto a Gold Sensor Surface

Abstract

Mammalian olfactory receptors (ORs) constitute the largest family of G-protein coupled receptors, with up to about 1000 different genes per species, each having specific odorant ligands. ORs could be used as sensing elements of highly specific and sensitive bioelectronic hybrid devices such as bioelectronic noses. After optimized immobilization onto the device, natural ORs provide molecular recognition of various odors with their intrinsic sensitivity, discrimination, and detection properties. However, the main difficulties are related to the low expression level of recombinant ORs, their stability and potential loss of activity. Such drawbacks can be successfully overcome in bioelectronic noses integrating nanosomes (nanometric membrane vesicles carrying ORs) that are stably immobilized through a specific antibody. The advantages of such a platform rely on the fact that ORs stay in the natural membrane environment and thus, preserve their full activity. Thanks to their small sizes, nanosomes offer potential for micro-and nano-scale sensor development. In this paper, we summarize the key elements regarding nanosomes production and manipulation and provide an example of their immobilization onto a gold sensor surface. Rat ORI7 is used as representative OR that can be functionally expressed in Saccharomyces cerevisiae. The receptor was not purified but only nanosomes were prepared. Nanosomes were immobilized onto functionalized gold surface using the anti-I7 antibody. Utilization of the antibody provides enrichment of ORI7 on the sensor surface but also uniform and appropriate orientation of the receptors. These features are crucial in optimization of bioelectronic nose' analytical performances.

Investigation of the Affinity of Aptamers for Bacteria by Surface Plasmon Resonance Imaging Using Nanosomes

“…Subsequent sonication of the preparation results in uniform vesicle sizes. 22 This simple protocol was applied to B. cereus, B. subtilis, and E. coli bacteria. The resulting nanosomes were characterized by DLS and NTA (Table 2).…”

“…A sonication of a bacterial suspension breaks the phospholipid membranes to form fragments, which spontaneously reassemble to form nanosized vesicles. Subsequent sonication of the preparation results in uniform vesicle sizes . This simple protocol was applied to B. cereus , B. subtilis , and E. coli bacteria.…”