Kinetics of Ligand Binding to Membrane Receptors from Equilibrium Fluctuation Analysis of Single Binding Events
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2212683
- author
- Gunnarsson, Anders ; Dexlin Mellby, Linda LU ; Wallin, Patric ; Svedhem, Sofia ; Jonsson, Peter LU ; Wingren, Christer LU and Hook, Fredrik
- organization
- publishing date
- 2011
- 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
- 2024-10-09 18:36:58
@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}}, }