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.

Original publication

DOI

10.1016/j.ab.2009.04.013

Type

Journal article

Journal

Analytical biochemistry

Publication Date

07/2009

Volume

390

Pages

203 - 205

Addresses

Manchester Interdisciplinary Biocenter, University of Manchester, Manchester M1 7DN, UK. mark.platt@manchester.ac.uk

Keywords

Humans, Thrombin, Protein Array Analysis, Surface Plasmon Resonance, Protein Binding, SELEX Aptamer Technique, Aptamers, Nucleotide