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

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

STAR protocols (2021-06-15)
Demetra S M Chatzileontiadou, Christopher Szeto, Dhilshan Jayasinghe, Stephanie Gras
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt dihydrate, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)
Sigma-Aldrich
Poly(ethylene glycol), BioUltra, 3,350
Sigma-Aldrich
L-Arginine monohydrochloride, reagent grade, ≥98% (HPLC), powder
Sigma-Aldrich
Bromophenol Blue, titration: suitable