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Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens
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Chapters
- 00:05Title
- 01:05T-cell Receptor (TCR) and Human Leukocyte Antigen-resticted Tumor-derived Peptide (pHLA) Refolding and Purification by Fast Protein Liquid Chromatography (FPLC)
- 03:43Surface Plasmon Resonance (SPR) Equilibrium Binding and Thermodynamic Analyses
- 05:55Protein Crystallization and X-ray Diffraction
- 07:39Results: Molecular Analysis of gp100280-288 Recognition by CD8+ T-cells
- 08:59Conclusion
Here, we describe methods that we commonly employ in the laboratory to determine how the nature of the interaction between the T-cell receptor and tumor antigens, presented by human leukocyte antigens, governs T-cell functionality; these methods include protein production, X-ray crystallography, biophysics, and functional T-cell experiments.
