In the search for cross-reactivity between bacteria and HLA-B27, three groups of investigators have identified several bacterial envelope proteins which are reactive with the monoclonal anti-HLA-B27 antibodies B27.M1 and B27.M2. Since these two antibodies react poorly with HLA-B27-derived synthetic peptides, it is not possible to locate the reactive epitopes on the HLA-B27 using synthetic peptides. Here, we introduce Ye-2, a monoclonal anti-HLA-B27 antibody which, unlike B27.M1 and B27.M2, is reactive with a synthetic peptide derived from residues 63-84 of HLA-B27.1. Analysis with a cross-reactive peptide derived from residues 226-244 of bovine carbonic anhydrase suggests that only a few of the amino acid residues in the HLA-B27-derived peptide are responsible for the reactivity. This antibody should be a useful adjunct in a preliminary assessment of whether a bacterial protein mimics HLA-B27 in primary structure.