title: Thermophoresis in nanoliter droplets to quantify aptamer binding.
abstract: 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.
authors.affiliation: Systems Biophysics, Physics Department, NanoSystems Initiative Munich and Center for Nanoscience, Ludwig-Maximilians-University Munich, Amalienstrasse 54, 80799 Munich (Germany) http://www.biosystems.physik.lmu.de.
authors.forename: Susanne A I
authors.initials: SA
authors.lastname: Seidel
authors.forename: Niklas A
authors.initials: NA
authors.lastname: Markwardt
authors.forename: Simon A
authors.initials: SA
authors.lastname: Lanzmich
authors.forename: Dieter
authors.initials: D
authors.lastname: Braun
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D002352:Carrier Proteins; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008968:Molecular Conformation
chemical_list: D052157:Aptamers, Nucleotide; D002352:Carrier Proteins
keywords: analytical methods; binding affinity; high-throughput screening; nanoliter thermophoresis; numerical simulation