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Abstract

We have developed an alternative Western Blot protocol based on AptaBodies replacing antibodies in the detection of proteins. AptaBodies consist of a DNA aptamer having a high affinity and specificity against its target molecule that is bound to a hemin/G-quadruplex horseradish peroxidase-mimicking DNAzyme (HRP DNAzyme). Already known aptamers or aptamers generated by our newly developed software MAWS were applied to build AptaBodies. As a proof of principle we applied AptaBodies to detect His-tagged proteins. Using the AptaBody-based approach, the same protocol as for conventional antibody-based Western blotting can be applied. Our innovative technology shows a variety of advantages. AptaBodies are a cheap, fast and a simple alternative to antibodies. Furthermore, time-consuming animal experiments to generate antibodies can be soon a thing of the past. Therefore, AptaBodies provide a promising alternative compared to antibody-based Western blotting.