Establishment of a quantitative ELISA capable of determining peptide - MHC class I interaction

Tissue Antigens. 2002 Apr;59(4):251-8. doi: 10.1034/j.1399-0039.2002.590402.x.


Many different assays for measuring peptide-MHC interactions have been suggested over the years. Yet, there is no generally accepted standard method available. We have recently generated preoxidized recombinant MHC class I molecules (MHC-I) which can be purified to homogeneity under denaturing conditions (i.e., in the absence of any contaminating peptides). Such denatured MHC-I molecules are functional equivalents of "empty molecules". When diluted into aqueous buffer containing beta-2 microglobulin (beta2m) and the appropriate peptide, they fold rapidly and efficiently in an entirely peptide dependent manner. Here, we exploit the availability of these molecules to generate a quantitative ELISA-based assay capable of measuring the affinity of the interaction between peptide and MHC-I. This assay is simple and sensitive, and one can easily envisage that the necessary reagents, standards and protocols could be made generally available to the scientific community.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Buffers
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Histocompatibility Antigens Class I / chemistry
  • Histocompatibility Antigens Class I / isolation & purification
  • Histocompatibility Antigens Class I / metabolism*
  • Humans
  • Peptides / chemistry
  • Peptides / immunology
  • Peptides / metabolism
  • Protein Binding / immunology
  • Protein Renaturation
  • Sensitivity and Specificity
  • beta 2-Microglobulin


  • Buffers
  • Histocompatibility Antigens Class I
  • Peptides
  • beta 2-Microglobulin