(352c) Cell-free synthesis goes electric: dual optical and electronic Biosensor via direct protein integration into a supported membrane Electrode

Transmembrane proteins (TMPs) are Nature’s biorecognition, sensing, and transduction
elements and critical components in biomimetic sensing. However, they are among the most
challenging biomolecules to integrate into biosensors because they require a lipid bilayer to
remain functional. Successful bilayer and TMP integration with bioelectronic devices would enable
new capabilities in in vitro biosensing and biological actuation. To date, reconstitution of lipid
bilayers onto sensing surfaces requires significant optimization of the abiotic/biotic interface.
Nonetheless, current hybrid biotic-abiotic devices show great promise in biosensing, but even
more so if arrays of TMPs tailored to sense specific targets could be produced. Such scale up is
necessary for rapidly identifying the most sensitive protein sensors, providing a myriad of sensing
elements that can monitor a variety of targets simultaneously. Our team has developed several
approaches to integrate TMPs into supported lipid bilayers that functionalize the surface of
organic bioelectronic devices. In this presentation, I will highlight an approach we pioneered using
cell-free synthesis to integrate TMPs directly into lipid bilayers that coat the sensing surface of a
planar electrode. We sense TMP properties, functions, and responses to chemical stimuli using
the dual modality of electronic and optical means. The criticality of dual mode data collection is
that, for transporters and ion channels, electrical means can be used label-free to read out flux of
material across the membrane. Optical approaches offer the possibility to simultaneously obtain
confirmation of folding with protein folding reporters, as well as obtaining orientation information.
Combining all these features into one platform offers a wealth of possibilities for many biosensing
and bioanalytical applications, including those that leverage TMPs for environmental sensing.