Aptamer evolution for array-based diagnostics |
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Authors: | Mark Platt William Rowe David C Wedge Douglas B Kell Joshua Knowles Philip JR Day |
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Institution: | aManchester Interdisciplinary Biocenter, University of Manchester, Manchester M1 7DN, UK;bSchool of Chemistry, University of Manchester, Manchester M13 9PL, UK;cSchool of Computer Science, University of Manchester, Manchester M13 9PL, UK;dSchool of Translational Medicine, University of Manchester, Manchester M13 9PT, UK |
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Abstract: | Closed loop aptameric directed evolution, (CLADE) is a technique enabling simultaneous discovery, evolution, and optimization of aptamers. It was previously demonstrated using a fluorescent protein, and here we extend its applicability with the generation of surface-bound aptamers for targets containing no natural fluorescence. Starting from a random population, in four generations CLADE produced a new aptamer to thrombin with high specificity and affinity. The best aptameric sequence was void of the set of four guanine repeats typifying thrombin aptamers and, thus, highlights the benefits of evolution performed in an environment closely mimicking the final diagnostic application. |
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