Aptamer evolution for array-based diagnostics.
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.
Platt
Mark
M
Manchester Interdisciplinary Biocenter, University of Manchester, Manchester M1 7DN, UK. mark.platt@manchester.ac.uk
Rowe
William
W
Wedge
David C
DC
Kell
Douglas B
DB
Knowles
Joshua
J
Day
Philip J R
PJ
D052157:Aptamers, Nucleotide / Q000138:chemical synthesis* / Q000378:metabolism; D006801:Humans; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D052156:SELEX Aptamer Technique / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000379:methods*; D013917:Thrombin / Q000032:analysis* / Q000378:metabolism
D052157:Aptamers, Nucleotide; C081676:thrombin aptamer; D013917:Thrombin