Abstract
The characterization and development of highly specific aptamers requires the analysis of the interaction strength between aptamer and target. MicroScale Thermophoresis (MST) is a rapid and precise method to quantify biomolecular interactions in solution at microliter scale. The basis of this technology is a physical effect referred to as thermophoresis, which describes the directed movement of molecules through temperature gradients. The thermophoretic properties of a molecule depend on its size, charge, and hydration shell. Since at least one of these parameters is altered upon binding of a ligand, this method can be used to analyze virtually any biomolecular interaction in any buffer or complex bioliquid. This section provides a detailed protocol describing how MST is used to obtain quantitative binding parameters for aptamer–target interactions. The two DNA-aptamers HD1 and HD22, which are targeted against human thrombin, are used as model systems to demonstrate a rapid and straightforward screening approach to determine optimal buffer conditions.
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Breitsprecher, D., Schlinck, N., Witte, D., Duhr, S., Baaske, P., Schubert, T. (2016). Aptamer Binding Studies Using MicroScale Thermophoresis. In: Mayer, G. (eds) Nucleic Acid Aptamers. Methods in Molecular Biology, vol 1380. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3197-2_8
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DOI: https://doi.org/10.1007/978-1-4939-3197-2_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3196-5
Online ISBN: 978-1-4939-3197-2
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