Empty MHC Class I Protein Production

This chapter contains a detailed protocol for the recombinant production and in vitro refolding of disulfide-stabilized peptide-empty MHC class I proteins. MHC class I proteins are heterotrimeric complexes consisting of a heavy chain, light chain (β₂m), and a peptide. This protocol covers MHC class I heavy chain and light chain expression in E. coli as inclusion bodies, their purification, and subsequent refolding using a dilution method. The protocol details the steps for expressing the proteins, purifying inclusion bodies, and refolding disulfide-stabilized MHC class I molecules with dipeptides. This approach ensures the production of correctly folded, functional, peptide-empty MHC class I proteins suitable for various biochemical and biophysical studies.