Research Article Single-Round Patterned DNA Library Microarray Aptamer Lead Identification

License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. A method for identifying an aptamer in a single round was developed using custom DNAmicroarrays containing computationally derived patterned libraries incorporating no information on the sequences of previously reported thrombin binding aptamers.The DNA library was specifically designed to increase the probability of binding by enhancing structural complexity in a sequence-space confined environment, much like generating lead compounds in a combinatorial drug screening library. The sequence demonstrating the highest fluorescence intensity upon target addition was confirmed to bind the target molecule thrombin with specificity by surface plasmon resonance, and a novel imino protonNMR/2DNOESY combination was used to screen the structure for G-quartet formation.We propose that the lack of G-quartet structure inmicroarray-derived aptamers may highlight differences in binding mechanisms between surface-immobilized and solution based strategies. This proof-of-principle study highlights the use of a computational driven methodology to create a DNA library rather than a SELEX based approach.This work is beneficial to the biosensor field where aptamers selected by solution based evolution have proven challenging to retain binding function whe