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Kinetics of Ligand Binding to Membrane Receptors from Equilibrium Fluctuation Analysis of Single Binding Events

Anders Gunnarsson (Institutionen för teknisk fysik, Biologisk fysik) ; L. Dexlin ; Patric Wallin (Institutionen för teknisk fysik, Biologisk fysik) ; Sofia Svedhem (Institutionen för teknisk fysik, Biologisk fysik) ; Peter Jönsson (Institutionen för teknisk fysik, Biologisk fysik) ; C. Wingren ; Fredrik Höök (Institutionen för teknisk fysik, Biologisk fysik)
Journal of the American Chemical Society (0002-7863). Vol. 133 (2011), 38, p. 14852-14855.
[Artikel, refereegranskad vetenskaplig]

Equilibrium fluctuation analysis of single binding events has been used to extract binding kinetics of ligand interactions with cell-membrane bound receptors. Time-dependent total internal reflection fluorescence (TIRF) imaging was used to extract residence-time statistics of fluorescently stained liposomes derived directly from cell membranes upon their binding to surface-immobilized antibody fragments. The dissociation rate constants for two pharmaceutical relevant antibodies directed against different B-cell expressed membrane proteins was clearly discriminated, and the affinity of the interaction could be determined by inhibiting the interaction with increasing concentrations of soluble antibodies. The single-molecule sensitivity made the analysis possible without overexpressed membrane proteins, which makes the assay attractive in early drug-screening applications.

Nyckelord: fluorescence correlation spectroscopy, target residence time, drug, discovery, mismatch discrimination, plasma-membranes, pharmacology, antibodies, proteomics, diversity, evolution



Denna post skapades 2011-11-03.
CPL Pubid: 148093

 

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Institutioner (Chalmers)

Institutionen för teknisk fysik, Biologisk fysik (2007-2015)

Ämnesområden

Kemi

Chalmers infrastruktur

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