Difference between revisions of "Template:Heidelberg/pages/ab/DiscussionAndOutlook"

In this project we show a new, efficient, and specific Western blot assay based on AptaBodies for the detection proteins. The fusion of a HRP DNAzyme with an aptamer, that binds to a POI, so called AptaBody is a promising alternative that could complement and sometimes even replace classical antibodies so far applied in Western Blot experiments.

As a proof of principle we targeted His-tag proteins using antiHis AptaBodies. Even in cell lysates the AptaBody is able to detected its target protein with a high specificity. Using AptaBodies instead of Antibodies may potentially have the following benefits:

1. The protocol is cost and time saving. The costs for an AptaBody are just those for an oligo DNA strand. Furthermore, no blocking step is needed to achieve a specific signal. (Fig. 12)
2. The generation of new AptaBodies is much faster than development of new antibodies. The design and production of the AptaBodies takes 14 days maximum. Please note that AptaBodies can be readily designed for proteins for which no antibodies are available yet.
3. MAWS can generate specific AptaBodies against each and every protein of interest.
4. In contrast to antibodies the production of AptaBodies do not require animal experiments.

Aptamers and methods for their in vitro selection and uses thereof. (o. J.). From http://www.google.com/patents/US20050142582

Dna aptamers binding the histidine tag and their application. (o. J.). From http://www.google.com/patents/WO2014185802A1

Wang, H., Wang, D. M., & Huang, C. Z. (2015). Highly sensitive chemiluminescent detection of lead ion based on its displacement of potassium in G-Quadruplex DNAzyme. The Analyst, 140(16), 5742–5747. http://doi.org/10.1039/c5an00884k