Kinetics of Ligand Binding to Membrane Receptors from Equilibrium Fluctuation Analysis of Single Binding Events
Artikel i vetenskaplig tidskrift, 2011

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.

mismatch discrimination

plasma-membranes

diversity

proteomics

drug

evolution

discovery

antibodies

pharmacology

fluorescence correlation spectroscopy

target residence time

Författare

Anders Gunnarsson

Chalmers, Teknisk fysik, Biologisk fysik

L. Dexlin

Lunds universitet

Patric Wallin

Chalmers, Teknisk fysik, Biologisk fysik

Sofia Svedhem

Chalmers, Teknisk fysik, Biologisk fysik

Peter Jönsson

Chalmers, Teknisk fysik, Biologisk fysik

C. Wingren

Lunds universitet

Fredrik Höök

Chalmers, Teknisk fysik, Biologisk fysik

Journal of the American Chemical Society

0002-7863 (ISSN) 1520-5126 (eISSN)

Vol. 133 38 14852-14855

Ämneskategorier

Kemi

DOI

10.1021/ja2047039