A new approach to synthetic chemistry is performed in ultraminiaturized, nanofabricated reaction chambers. Using lithographically defined nanowells, we achieve single-point covalent chemistry on hundreds of individual carbon nanotube transistors, providing robust statistics and unprecedented spatial resolution in adduct position. Each device acts as a sensor to detect, in real-time and through quantized changes in conductance, single-point functionalization of the nanotube as well as consecutive chemical reactions, molecular interactions, and molecular conformational changes occurring on the resulting single-molecule probe. In particular, we use a set of sequential bioconjugation reactions to tether a single-strand of DNA to the device and record its repeated, reversible folding into a G-quadruplex structure. The stable covalent tether allows us to measure the same molecule in different solutions, revealing the characteristic increased stability of the G-quadruplex structure in the presence of potassium ions (K+) versus sodium ions (Na+). Nanowell-confined reaction chemistry on carbon nanotube devices offers a versatile method to isolate and monitor individual molecules during successive chemical reactions over an extended period of time.

The study of biomolecular interactions at the single-molecule level holds great potential for both basic science and biotechnology applications. Single-molecule studies often rely on fluorescence-based reporting, with signal levels limited by photon emission from single optical reporters. The point-functionalized carbon nanotube transistor, known as the single-molecule field-effect transistor, is a bioelectronics alternative based on intrinsic molecular charge that offers significantly higher signal levels for detection. Such devices are effective for characterizing DNA hybridization kinetics and thermodynamics and enabling emerging applications in genomic identification. In this work, we show that hybridization kinetics can be directly controlled by electrostatic bias applied between the device and the surrounding electrolyte. We perform the first single-molecule experiments demonstrating the use of electrostatics to control molecular binding. Using bias as a proxy for temperature, we demonstrate the feasibility of detecting various concentrations of 20-nt target sequences from the Ebolavirus nucleoprotein gene in a constant-temperature environment.

Carbon-nanotube field-effect transistors (CNTFETs) have been used to sense conformational changes and binding events in protein and nucleic acid structures from the intrinsic molecular charge. The key to utilizing these CNT devices as single-molecule sensors is the ability to attach a single probe molecule to an individual device. In contrast with noncovalent attachment approaches such as those based on van der Waals interactions, covalent attachment approaches generally deliver higher stability but have traditionally been more difficult to control, resulting in low yield. Here we present a single-point-functionalization method for CNTFET arrays based on electrochemical control of a diazonium reaction to create sp3 defects, combined with a scalable spin-casting method for large arrays of devices on arbitrary substrates. Attachment of probe DNA to the functionalized device verifies correct single molecule detection of DNA hybridization with the complementary target. This method enables rapid single-point defect generation with 80% yield, rendering it adaptable to fabricating large arrays of CNTFET devices.

The olfactory receptor neurons of insects and vertebrates are gated by odorant receptor (OR) proteins of which several members have been shown to exhibit remarkable sensitivity and selectivity towards volatile organic compounds of significant importance in the fields of medicine, agriculture and public health. Insect ORs offer intrinsic amplification where a single binding event is transduced into a measurable ionic current. Consequently, insect ORs have great potential as biorecognition elements in many sensor configurations. However, integrating these sensing components onto electronic transducers for the development of biosensors has been marginal due to several drawbacks, including their lipophilic nature, signal transduction mechanism and the limited number of known cognate receptor-ligand pairs. We review the current state of research in this emerging field and highlight the use of a group of indole-sensitive ORs (indolORs) from unexpected sources for the development of biosensors.

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