Competition against allele-specific AMCA-labeled binding peptides.

Competition of synthetic HNP-2 (A) and isolated HNP from HL-60 cells (B) to solubilized HLA-DRB1*0101 (○), HLA-DRB1* HLA-DRB1*1501/DRB5*0101 (▴), HLA-DRB1*0301 (×), HLA-DRB1*0401 (▪) and HLA-DRB1*0701 (□) against fluorescently (AMCA)-labeled binding peptides using a gel filtration assay. As binding peptides, HA 306-18 (PKYVKQNTLKLAT; 1.5 μmol/L) was used for HLA-DR1, -2 and -4. For HLA-DR3, heat shock protein 65, 3-13 (KTIAYDEEARR; 1.5 μmol/L) and for HLA-DR7, heat shock protein 70, 38-52 (TPSYVAFTDTERLIG; 1.5 μmol/L) was used. For (A), the concentration unit μg/40 μL can be transferred to μmol/L by multiplication with the factor 7.4. For (B), however, the μmol/L concentration of the used defensin isolate cannot be determined due to inhomogeneous masses of different defensin molecules.