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Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens
JoVE Journal
Imunologia e Infecção
This content is Free Access.
JoVE Journal Imunologia e Infecção
Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens
DOI:

09:53 min

February 06, 2017

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Capítulos

  • 00:05Título
  • 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

Summary

Tadução automática

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.

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