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Protein purification and crystallization of HLA-A∗02:01 in complex with SARS-CoV-2 peptides

STAR Protoc. 2021-06; 
Demetra S M Chatzileontiadou, Christopher Szeto, Dhilshan Jayasinghe, Stephanie Gras
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PCR Cloning and Subcloning … of HLA-A*02:01 heavy chain and human β2-microglobulin proteins as well as their recovery from inclusion bodies from Escherichia coli expression using a mild … Both sequences and successful sub-cloning into pET30a vector have been confirmed by sequencing by Genscript … Get A Quote

摘要

Understanding T-cell responses requires identifying viral peptides presented by human leukocyte antigens (HLAs). X-ray crystallography can be used to visualize their presentation. This protocol describes the expression, purification, and crystallization of HLA-A∗02:01, one of the most frequent HLA in the global population in complex with peptides derived from the SARS-CoV-2 nucleocapsid protein. This protocol can be applied to different HLA class I molecules bound to other peptides. For complete details on the use and execution of this protocol, please refer to Szeto et al. (2021).

关键词

Immunology, Protein expression and purification, X-ray Crystallography