Regensburg 2013 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 17: Biomaterials and Biopolymers II (joint with CPP)
BP 17.7: Vortrag
Mittwoch, 13. März 2013, 11:30–11:45, H43
FACS-sorting of particles to reduce the data variance in Optical Tweezers assisted Dynamic Force Spectroscopy measurements — •Tim Stangner1, David Singer2, Carolin Wagner1, Christof Gutsche1, Olaf Ueberschär1, Ralf Hoffmann2, and Friedrich Kremer1 — 1Universität Leipzig, Institut für Experimentelle Physik I, Linnéstraße 5, 04103 Leipzig, Deutschland — 2Biotechnologisch-Biomedizinisches Zentrum Leipzig, Fakultät für Chemie und Mineralogie, Deutscher Platz 5, 04103 Leipzig, Deutschland
By combining Optical Tweezers assisted dynamic force spectroscopy experiments with fluorescence activated cell sorting (FACS), we demonstrate a new approach to reduce the data variance in measuring receptor-ligand-interactions on a single molecule level by ensuring similar coating densities. Therefore, the carboxyfluorescein-labeled monophosphorylated peptide tau226-240[pThr231] is anchored on melamine resin beads and these beads are sorted by FACS to achieve a homogeneous surface coverage. To quantify the impact of the fluorescence dye on the bond parameters between the phosphorylated peptide and the corresponding phosphorylation specific anti-human tau monoclonal antibody HPT-104, we perform dynamic force spectroscopy and compare the results to data using unsorted beads covered with the non-fluorescence peptide analogue. Finally, we demonstrate that the data variance of the relative binding frequency is significantly decreased by a factor of 3.4 using presorted colloids with a homogeneous ligand coating compared to unsorted ones.