TY - JOUR

T1 - Establishment of a quantitative ELISA capable of determining peptide - MHC class I interaction.

AU - Sylvester-Hvid, C

AU - Kristensen, N

AU - Blicher, T

AU - Ferré, H

AU - Lauemøller, S L

AU - Wolf, X A

AU - Lamberth, K

AU - Nissen, Mogens Holst

AU - Pedersen, L Ø

AU - Buus, S

N1 - Keywords: Buffers; Enzyme-Linked Immunosorbent Assay; Histocompatibility Antigens Class I; Humans; Peptides; Protein Binding; Protein Renaturation; Sensitivity and Specificity; beta 2-Microglobulin

PY - 2002

Y1 - 2002

N2 - 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.

AB - 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.

M3 - Journal article

C2 - 12135423

VL - 59

SP - 251

EP - 258

JO - HLA

JF - HLA

SN - 2059-2302

IS - 4

ER -