characterization strategy will be presented. Finally, issues of comparability of
binding data between different aptamer characterization technologies will be
discussed.
Graphical Abstract
Keywords Affinity, Binding parameters, Biolayer Interferometry, Biophysical
characterization, EMSA, Filter-Binding Assay, Flow Cytometry, Fluorescence
Polarization, Isothermal Titration Calorimetry, Kinetics, MicroScale
Thermophoresis, Surface Plasmon Resonance, Thermodynamics, SwitchSense
1 Basic Binding Parameters in Aptamer Development
Basic biophysical binding parameters such as affinity, kinetics, or thermodynamics
are key aspects in the development of aptamers for pharmaceutical and diagnostic
use. Binding affinity is a measure of binding strength between aptamer and target
and is usually reported as an equilibrium dissociation constant (K D ). The lower this
constant, the higher the binding strength between aptamer and target. In the course of
aptamer development, affinity is often used to initially rank a pool of aptamers to
select relevant candidates. In addition, affinity enables to express selectivity of
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M. Plach and T. Schubert