Dungl, D. (2019). Binding kinetics between the T cell receptor and its ligand pMHC [Diploma Thesis, Technische Universität Wien]. reposiTUm. https://doi.org/10.34726/hss.2019.66563
single molecule microscopy; FRET; T cell; immunological synapse; off-rate
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Abstract:
The recognition of pathogens by the T cell receptor plays a crucial role in the immune system. The binding kinetics between the T cell receptor and its ligand are still debated. They have been characterized by Huppa et al. using a fluorescence microscopy combined with a Förster Resonant Energy Transfer (FRET) based approach and reported off times of approximately 0.1 s. In contrast ODonoghue et al. reported different off times of approximately 5 s using fluorescence microscopy with long exposure times. In this thesis, it was attempted to reconcile both results by combining the methods of both papers to measure the binding kinetics of T cell receptors and its ligands. Experimental procedures were established and receptorligand interactions were observed using long exposures, but the FRET experiments were inconclusive due to similar results in negative control experiments.
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