Label-free monitoring of DNA-ligand interactions

Analytical Biochemistry
J PiehlerS Grabley

Abstract

We report on the label- and isotope-free monitoring of DNA interactions with low-molecular-weight ligands. An optical technique based on interference at thin layers was used to monitor in real time binding of ligands at DNA which was immobilized by Coulomb interactions at a positively charged surface. Approximately 2 ng DNA/m2 was irreversibly bound to the surface, which remained stable over several days. This result was confirmed by characterization of the layer using spectroscopic ellipsometry. During incubation of immobilized DNA with a variety of intercalators and other DNA-binding compounds in a flow system, interactions were monitored by reflectometric interference spectroscopy. Binding effects between 10 and 400 pg/ mm2 were detected unambiguously. Nonspecific binding effects were excluded by using a negatively charged reference surface. Variation of intercalator concentration allowed the characterization of interaction with respect to kinetics and thermodynamics by the evaluation of binding rate and equilibrium coverage. The affinity constants were determined in the range between 10(5) and 10(6) M-1, in good agreement to those obtained by homogeneous phase assays. Association rate constants between 10(3) and 10(5) M-1 s...Continue Reading

Citations

Jan 10, 2002·Analytical Biochemistry·Hans-Martin HaakeGünter Gauglitz
Feb 22, 2012·Analytical and Bioanalytical Chemistry·Urs HilbigGünter Gauglitz
Jul 16, 2014·Journal of Ethnopharmacology·Anna Rita Bilia
Sep 4, 2009·Analytical and Bioanalytical Chemistry·Thomas SchülerJürgen Popp
Jul 14, 2010·Analytical and Bioanalytical Chemistry·Günter Gauglitz
Apr 27, 2007·Journal of the American Society for Mass Spectrometry·Frédéric RosuValérie Gabelica
Apr 3, 2013·Current Protocols in Protein Science·F X Reymond SutandyHeng Zhu
Oct 28, 2017·The Journal of Biological Chemistry·Birte PlitzkoSandra Loesgen
Sep 5, 2001·Nature Biotechnology·G WuA Majumdar
Aug 29, 2006·Angewandte Chemie·Sebastian J KochThomas Schrader
Apr 30, 2009·Biopolymers·V V KostjukovM P Evstigneev
Nov 19, 2008·Nanotechnology·Ning ShaoBalaji Panchapakesan
Feb 8, 2019·Chembiochem : a European Journal of Chemical Biology·Normann KilbGünter Roth
Nov 26, 2020·Analytical and Bioanalytical Chemistry·Ross D OverackerSandra Loesgen
Apr 21, 2021·Journal of Nanoscience and Nanotechnology·Ao DongWeiping Qian
Jan 13, 2006·Journal of the American Chemical Society·Christian RennerThomas Schrader
Oct 12, 2000·Analytical Chemistry·D W Pang, H D Abruña
Dec 9, 1998·Analytical Chemistry·A B SteelM J Tarlov

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