JoVE Journal

Immunology and Infection

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利用X射线晶体学，生物物理学，和功能分析，以确定管理机制T细胞癌抗原的受体识别

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11.4K Views

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09:53 min

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February 6th, 2017

DOI :

10.3791/54991-v

February 6th, 2017

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Bruce J. MacLachlan*¹, Alexander Greenshields-Watson*¹, Georgina H Mason*¹, Andrea J Schauenburg¹, Valentina Bianchi¹^,²^,³, Pierre J Rizkallah¹, Andrew K Sewell¹, Anna Fuller¹, David K Cole¹

¹Division of Infection and Immunity and Systems Immunity Research Institute, Cardiff University, ²Department of Oncology, University Hospital of Lausanne (CHUV), ³Ludwig Insitutue for Cancer Research, Lausanne Branch, University of Lausanne

摘要

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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120 CD8 T T TCR PHLA X gp100 heteroclitic

此视频中的章节

0:05

Title

1:05

T-cell Receptor (TCR) and Human Leukocyte Antigen-resticted Tumor-derived Peptide (pHLA) Refolding and Purification by Fast Protein Liquid Chromatography (FPLC)

3:43

Surface Plasmon Resonance (SPR) Equilibrium Binding and Thermodynamic Analyses

5:55

Protein Crystallization and X-ray Diffraction

7:39

Results: Molecular Analysis of gp100_280-288 Recognition by CD8⁺ T-cells

8:59

Conclusion

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