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

Gunnarsson, Anders ; Dexlin Mellby, Linda LU ; Wallin, Patric ; Svedhem, Sofia ; Jonsson, Peter LU ; Wingren, Christer LU and Hook, Fredrik (2011) In Journal of the American Chemical Society 133(38). p.14852-14855
Abstract
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... (More)
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. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of the American Chemical Society
volume
133
issue
38
pages
14852 - 14855
publisher
The American Chemical Society (ACS)
external identifiers
  • wos:000295604400004
  • scopus:80053084242
  • pmid:21866974
ISSN
1520-5126
DOI
10.1021/ja2047039
language
English
LU publication?
yes
id
0026c260-0e9c-4b6f-927e-87400bf4d81f (old id 2212683)
date added to LUP
2016-04-01 12:54:07
date last changed
2022-04-21 18:33:15
@article{0026c260-0e9c-4b6f-927e-87400bf4d81f,
  abstract     = {{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.}},
  author       = {{Gunnarsson, Anders and Dexlin Mellby, Linda and Wallin, Patric and Svedhem, Sofia and Jonsson, Peter and Wingren, Christer and Hook, Fredrik}},
  issn         = {{1520-5126}},
  language     = {{eng}},
  number       = {{38}},
  pages        = {{14852--14855}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Journal of the American Chemical Society}},
  title        = {{Kinetics of Ligand Binding to Membrane Receptors from Equilibrium Fluctuation Analysis of Single Binding Events}},
  url          = {{http://dx.doi.org/10.1021/ja2047039}},
  doi          = {{10.1021/ja2047039}},
  volume       = {{133}},
  year         = {{2011}},
}