An assay for simultaneous multiple determinations of peptide binding to MHC class II molecules.
Binding of positively charged radiolabeled synthetic peptides to human major histocompatibility complex class II DR molecules, purified by affinity chromatography from lymphoblastoid B cell lines of different haplotypes, is rapidly, quantitatively, and specifically assayed by selective adsorption of the complexes between peptide and DR molecules onto DEAE-cellulose paper disks. This assay can be used as a revealing system of the ability of unlabeled test peptides to competitively inhibit the binding between the radiolabeled peptide and the DR molecules, thus measuring the binding of the competitor peptides, irrespective of their charge properties, to different DR molecules.
Manfredi
A A
AA
Department of Biochemistry, CBS, University of Minnesota, St. Paul 55108.
Yuen
M H
MH
Raftery
M A
MA
Conti-Tronconi
B M
BM
D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000737:chemistry; D001667:Binding, Competitive; D002460:Cell Line; D002846:Chromatography, Affinity; D006684:HLA-DR Antigens / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D006801:Humans; D007457:Iodine Radioisotopes; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D007457:Iodine Radioisotopes; D010455:Peptides