Covalent assembly of a soluble T cell receptor-peptide-major histocompatibility class I complex

We used stepwise photochemical crosslinking for specifically assembling soluble and covalent complexes made of a T-cell antigen receptor (TCR) and a class I molecule of the major histocompatibility complex (MHC) bound to an antigenic peptide, For that purpose, rye have produced in myeloma cells a single-chain Fv construct of a TCR specific for a photoreactive H-2K(d)-peptide complex. Photochemical cross-linking of this TCR single-chain Fv with a soluble form of the photoreactive H-2K(d)-peptide ligand resulted in the formation of a ternary covalent complex. We have characterized the soluble ternary complex and shoved that it reacted with antibodies specific for epitopes located either on the native TCR or on the K-d molecules, By preventing the fast dissociation kinetics observed with most T cell receptors, this approach provides a means of preparing soluble TCR-peptide-MHC complexes on large-scale levels.