"Fitting" Makes "Sensing" Simple: Label-Free Detection Strategies Based on Nucleic Acid Aptamers

Nucleic acid aptamers are small sequences of DNA made via in vitro selection techniques to bind targets with high affinity and specificity. The term aptamer derives from the Latin, aptus, meaning "to fit", emphasizing the lock-and-key relationship between aptamers and their binding targets. In 2004, aptamers began to attract researchers attention as new binding elements for biosensors (i.e. aptasensors). Their advantages over other sensors include a diverse range of possible target molecules, high target affinity, simple synthesis, and ability to form Watson-Crick base pairs. These attributes create an enormous array of possible sensing applications and target molecules, spanning nearly all detection methods and readout techniques. In particular, aptamers provide an opportunity for designing "label-free" sensors, meaning sensors that do not require covalently labeling a signal probe to either the analyte or the recognition element (here, the aptamer). "Label-free" systems previously could only analyze large molecules using a few readout techniques, such as when employing the other recognition elements like antibodies. "Label-free" methods are one of the most effective and promising strategies for faster, simpler, and more convenient detection, since they avoid the expensive and tedious labeling process and challenging labeling reactions, while retaining the highest degree of activity and affinity for the recognition element. "Label-free" sensors are one of the most promising future biosensors. In this Account, we describe our efforts exploring and constructing such label-free sensing strategies based on aptamers. Our methods have included using various readout techniques, employing novel nanomaterials, importing lab-on-a-chip platforms, and improving logical recognition. The resulting sensors demonstrate that aptamers are ideal tools for "label-free" sensors. We divide this Account into three main parts describing three strategies for designing label-free sensors: (1) Label-free, separation-free strategies. These include colorimetric sensors based on G-quadruplex-hemin complex, and fluorescent sensors based on fluorescent small molecules, novel conjugated polymers, and metal ion clusters. (2) Label-free, separation-required strategies. In this part, electrochemical sensors are introduced, including sensors with different subtechniques using an electrode array. (3) Logic sensors. Some logic recognition systems are introduced. We emphasize that label-free aptasensors are not merely simple. We hope our introduction illustrates the powerful, flexible, and smart functions of aptamers in carrying out various detection tasks or playing various recognition games. Our work is only a start. We believe this field will bring additional knowledge on general designs, anti-interference, multianalysis, minimization, and auto-operation of aptamer biosensors.