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DTIC ADA551279: Ready-to-use Aptamer Biosensors for DNT and RDX

Aptamers are nucleic acid-based reagents that bind to target molecules with high affinity and specificity. However, methods for generating aptamers from random combinatorial libraries (e.g., SELEX) are often labor-intensive and time-consuming. To address this problem, we have recently demonstrated the microfluidic SELEX (M-SELEX) technology that can accelerate aptamer isolation by enabling highly stringent selection conditions through the use of very small amounts of target molecules (X. Lou et a/, PNAS 2009). In this work, we utilize the M-SELEX technology to generate a novel class of Self-Reporting Aptamers (SRAs) which are capable of a) specifically bind to their target, b) undergo conformational change triggered by this binding event, and c) activate a DNA enzyme moiety within the aptamer structure to fluorescently report the binding event. This technique is especially useful for the detection of small molecules because it does not require immobilization of the target molecules on solid support. As a proof of concept, in the last reporting period, we have demonstrated the isolation of SRAs that specifically bind to a target protein with low nanomolar affinity (S. Oh eta/., in r

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