Measuring interactions of MHC class I molecules using surface plasmon resonance

Sergei N. Khilko, Marie T. Jelonek, Maripat Corr, Lisa F. Boyd, Alfred L.M. Bothwell, David H. Margulies

Research output: Contribution to journalArticlepeer-review

51 Scopus citations


To examine the molecular interactions between major histocompatibility complex (MHC)-encoded molecules and peptides, monoclonal antibodies (mAbs), or T cell receptors, we have developed model systems employing genetically engineered soluble MHC class I molecules (MHC-I), synthetic peptides, purified mAbs, and engineered solubilizable T cell receptors. Direct binding assays based on immobilization of one of the interacting components to the dextran modified gold biosensor surface of a surface plasmon resonance (SPR) detector have been developed for each of these systems. The peptide binding site of the MHC-I molecule can be sterically mapped by evaluation of a set of peptides immobilized through the thiol group of cysteine substitutions at each peptide position. Kinetic binding studies indicate that the MHC-I/peptide interaction is characterized by a low to moderate apparent kass (∼ 5000-60000 M-1 s-1) and very small kdis (∼ 10-4-10-6 s-1) consistent with the biological requirement for a long cell surface residence time to permit engagement with T cell receptors. Several mAb directed against different MHC-I epitopes were examined, and kinetic parameters of their interaction with MHC molecules were determined. These showed characteristic moderate association rate constants and moderate dissociation rate constants (kass ∼ 104-106 M-1 s-1 and kdis ∼ 10-2-10-4 s-1), characteristic of many antibody/protein antigen interactions. The interaction of an anti-idiotypic anti-TCR mAb with its purified cognate TCR was of moderate affinity and revealed kinetic binding similar to that of the anti-MHC mAbs. The previously determined interaction of a purified T cell receptor with its MHC-I/peptide ligand is characterized by kinetic constants more similar to those of the antibody/antigen interaction than of the MHC-I/peptide interaction, but is remarkable for rapid dissociation rates (apparent kdis ∼ 10-2 s-1). Such binding studies of reactions involving the MHC-I molecules offer insight into the mechanisms responsible for the initial specific events required for the stimulation of T cells.

Original languageEnglish (US)
Pages (from-to)77-94
Number of pages18
JournalJournal of Immunological Methods
Issue number1
StatePublished - Jun 14 1995
Externally publishedYes


  • Major histocompatibility complex
  • Monoclonal antibody
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


Dive into the research topics of 'Measuring interactions of MHC class I molecules using surface plasmon resonance'. Together they form a unique fingerprint.

Cite this