Production and purification of infectious diseases-associated tetramer for staining of CD8(+) T cells from PBMCs

Major histocompatibility complex (MHC) tetramers can work as diagnostic tools to identify antigen-specific T cells in immunological research and monitoring. Here, we provide a general protocol for the production of MHC tetramer. We obtain highly pure N-terminal His-tagged HLA-A2 α chain and β2-micro...

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Detalles Bibliográficos
Autores principales: Ye, Jiezhou, Lei, Wen, Xiao, ChanChan, Su, Xiangmeng, Gao, Jie, Li, Xinran, Li, Wenxin, Wang, Pengcheng, Jiang, Zhenyou, Chen, Guobing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914994/
https://www.ncbi.nlm.nih.gov/pubmed/35284834
http://dx.doi.org/10.1016/j.xpro.2022.101206
Descripción
Sumario:Major histocompatibility complex (MHC) tetramers can work as diagnostic tools to identify antigen-specific T cells in immunological research and monitoring. Here, we provide a general protocol for the production of MHC tetramer. We obtain highly pure N-terminal His-tagged HLA-A2 α chain and β2-microglobulin (β(2)m) to fold a monomer with a photocleavable peptide, which can exchange with an HLA-A2 presented peptide derived from influenza A virus. Further those monomers compose tetramer to stain antigen-specific CD8(+) T cells. For complete details on the use and execution of this protocol, please refer to Xiao C.C. et al. (2021).

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