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Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface
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Chapters
- 00:05Title
- 01:35Red Blood Cell Biotinylation
- 03:05Functionalizing Biotin-Linked Ligands on Red Blood Cells
- 03:49Preparation of Micropipettes and Cell Chamber
- 04:51Micropipette Adhesion Frequency Assay
- 07:49Representative Results
- 12:27Conclusion
An adhesion frequency assay for measuring receptor-ligand interaction kinetics when both molecules are anchored on the surfaces of the interacting cells is described. This mechanically-based assay is exemplified using a micropipette-pressurized human red blood cell as adhesion sensor and integrin αLβ2 and intercellular adhesion molecule-1 as interacting receptors and ligands.
Tags
Adhesion Frequency AssayIn Situ Kinetics AnalysisCross-junctional Molecular InteractionsCell-cell InterfaceMicropipette Adhesion AssayReceptor-ligand Binding KineticsHuman Red Blood Cell (RBC)MicromanipulationBond FormationAdhesion EventRBC ElongationLigands ImmobilizedAdhesion ProbabilityBinding CurveProbabilistic ModelReceptor-ligand Reaction Kinetics2D AffinityOff-rateFcγ ReceptorsIgG Fc1-6SelectinsGlycoconjugate LigandsIntegrinsHomotypical Cadherin BindingT Cell ReceptorCoreceptorPeptide-major Histocompatibility Complexes
