Open AccessA New Spectral Shift-Based Method to Characterize Molecular Interactions
Author(s) -
Andreas Langer,
Tanja Bartoschik,
Ondrej Cehlár,
Stefan Duhr,
Burkart Philipp,
Werner Streicher
Publication year - 2022
Publication title -
assay and drug development technologies
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.402
H-Index - 56
eISSN - 1557-8127
pISSN - 1540-658X
DOI - 10.1089/adt.2021.133
Subject(s) - biomolecule , characterization (materials science) , chemistry , small molecule , ligand (biochemistry) , molecule , fluorescence , nucleic acid , molecular dynamics , drug discovery , biological system , biophysics , nanotechnology , materials science , computational chemistry , biology , biochemistry , physics , receptor , organic chemistry , quantum mechanics
There are many fluorescence-based applications that can be used to characterize molecular interactions. However, available methods often depend on site-specific labeling techniques or binding-induced changes in conformation or size of the probed target molecule. To overcome these limitations, we applied a ratiometric dual-emission approach that quantifies ligand-induced spectral shifts with sub-nanometer sensitivity. The use of environment-sensitive near-infrared dyes with the method we describe enables affinity measurements and thermodynamic characterization without the explicit need for site-specific labeling or ligand-induced conformational changes. We demonstrate that in-solution spectral shift measurements enable precise characterization of molecular interactions for a variety of biomolecules, including proteins, antibodies, and nucleic acids. Thereby, the described method is not limited to a subset of molecules since even the most challenging samples of research and drug discovery projects like membrane proteins and intrinsically disordered proteins can be analyzed.