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Thermophoresis in Nanoliter Droplets to Quantify Aptamer Binding
Author(s) -
Seidel Susanne A. I.,
Markwardt Niklas A.,
Lanzmich Simon A.,
Braun Dieter
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201402514
Subject(s) - microscale thermophoresis , thermophoresis , marangoni effect , miniaturization , biomolecule , microfluidics , bioanalysis , nanotechnology , capillary action , chemistry , aptamer , analytical chemistry (journal) , microtiter plate , materials science , chemical physics , chromatography , nanoparticle , convection , mechanics , nanofluid , biochemistry , physics , biology , composite material , genetics
Biomolecule interactions are central to pharmacology and diagnostics. These interactions can be quantified by thermophoresis, the directed molecule movement along a temperature gradient. It is sensitive to binding induced changes in size, charge, or conformation. Established capillary measurements require at least 0.5 μL per sample. We cut down sample consumption by a factor of 50, using 10 nL droplets produced with acoustic droplet robotics (Labcyte). Droplets were stabilized in an oil–surfactant mix and locally heated with an IR laser. Temperature increase, Marangoni flow, and concentration distribution were analyzed by fluorescence microscopy and numerical simulation. In 10 nL droplets, we quantified AMP‐aptamer affinity, cooperativity, and buffer dependence. Miniaturization and the 1536‐well plate format make the method high‐throughput and automation friendly. This promotes innovative applications for diagnostic assays in human serum or label‐free drug discovery screening.