The sequence encoding HLA‐B*18:01 heavy chain was obtained from the IPD‐IMGT/HLA database.
14 (link) The cDNA construct for the soluble protein insert, excluding the transmembrane domain, was sub‐cloned into the pET30 plasmid vector using NedI/HindIII restriction enzymes (GenScript, Piscataway, USA). The recombinant protein and human β2‐microglobulin (β2m) were then individually expressed in BL21‐RIL Escherichia coli cells, resulting in inclusion bodies that were extracted and purified through centrifugation.
49 (link) The peptides described in this study were chemically synthesised (GenScript; Tables1 and 2 ). Refolding was performed using 5 mg of peptide, 30 mg of HLA‐B*18:01 heavy chain and 10 mg of β2m in refolding buffer [3 m Urea (ThermoFisher, Scoresby, VIC, Australia), 0.4 m l ‐Arginine, (Merck, Darmstadt, Germany) 0.1 m Tris–HCl pH 8 (ThermoFisher), 2 mm Na‐EDTA pH 8 (Merck), 0.16% w/v reduced Glutathione (Goldbio, St Louis, USA) and 0.03% w/v oxidised Glutathione (Goldbio)]. This solution was dialyzed in 10 mM Tris–HCl pH 8 (ThermoFisher) and the peptide‐HLA‐B*18:01 complexes were purified using a two‐stage anion exchange chromatography (Cytiva, Marlborough, Massachusetts, USA).
14 (link) The cDNA construct for the soluble protein insert, excluding the transmembrane domain, was sub‐cloned into the pET30 plasmid vector using NedI/HindIII restriction enzymes (GenScript, Piscataway, USA). The recombinant protein and human β2‐microglobulin (β2m) were then individually expressed in BL21‐RIL Escherichia coli cells, resulting in inclusion bodies that were extracted and purified through centrifugation.
49 (link) The peptides described in this study were chemically synthesised (GenScript; Tables
