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A novel gastric‐cancer‐associated mucin antigen defined by a monoclonal antibody A3D4

De‐glycosylation of mucins may expose new tumor‐associated core protein epitopes. In this study, to attempt to develop useful markers for gastric cancers, we have purified and de‐glycosylated gastric mucin and tried to establish monoclonal antibodies (MAbs). A MAb designated A3D4 among established MAbs was shown to react with gastric cancer with high frequency, but not with normal gastric epithelium. Among normal digestive organs, only the colon and gall bladder were positive for MAb A3D4. The incidence of positivity in gastric cancer was 75% for intestinal‐type adenocarcinoma (n = 28), 40% for solid‐type adenocarcinoma (n = 5) and 33% for signet/scirrhous‐type adenocarcinoma (n = 15). Interestingly, adenoma and intestinal metaplasia (IM) with chronic gastritis or peptic ulcer were negative for MAb A3D4, whereas 8 out of 13 cases (62%) of IM with gastric cancer was positive. Western‐blot analysis using the lysate from normal colon tissues revealed a high‐molecular‐weight (>300‐kDa) smear‐like band. Immunohistochemical analysis indicated that the reactivity of MAb A3D4 was clearly increased when tissue sections were pre‐treated with periodic acid or O‐glycanase, while it was decreased by pre‐treatment with trypsin or protease V8. There was no reactivity with the synthetic peptide encompassing the tandem‐repeat sequence of MUC2 or MUC3. These data suggest that MAb A3D4 detects a novel gastric‐cancer‐associated mucin antigen whose epitope may be peptide in nature. Int. J. Cancer 73:795–801, 1997. © 1997 Wiley‐Liss, Inc.