Proto-Antigen-Presenting Synthetic Surfaces, Activated T Cells, and Uses Thereof

A method of forming a proto-antigen-presenting surface, the method comprising:

• synthesizing a plurality of major histocompatibility complex (MHC) molecules in the presence of initial peptide, thereby forming a plurality of complexes each comprising an MHC molecule and an initial peptide;
• wherein:
• the initial peptide is non-immunogenic;
• and (i) a plurality of primary activating molecules comprise the MHC molecules and first reactive moieties, or (ii) a plurality of primary activating molecules is prepared by adding first reactive moieties to the MHC molecules, and
• the method further comprises reacting the first reactive moieties of the plurality of primary activating molecules with a first plurality of binding moieties disposed on a covalently functionalized synthetic surface, thereby forming the proto-antigen-presenting surface.

A method of analyzing stability of a complex comprising a major histocompatibility complex (MHC) molecule and a peptide antigen, wherein the MHC molecule is configured to bind to a T cell receptor (TCR), the method comprising:

• contacting a plurality of the MHC molecules with the peptide antigen and an exchange factor, thereby forming peptide antigen-bound MHC molecules, wherein an initial peptide is bound to the MHC molecules before contact with the peptide antigen and exchange factor;
• wherein
• (i) a plurality of primary activating molecules comprise the MHC molecules and first reactive moieties or (ii) a plurality of primary activating molecules is prepared by adding first reactive moieties to the MHC molecules, and the method further comprises reacting the first reactive moieties of the plurality of primary activating molecules with a first plurality of binding moieties disposed on a covalently functionalized synthetic surface;
• and
• measuring total binding and/or an extent of dissociation of the peptide antigen from the MHC molecule.