Analysis of monoclonal antibodies reactive with meconium- and amniotic fluid-derived mucin.
The present study was undertaken to determine whether monoclonal antibodies (moABs TKH-2, MA54, MA61, B72.3, and CC49) directed toward O-linked mucin-type glycoprotein detect the NeuAc alpha 2-6GalNAc (sialyl Tn) epitope in meconium- and amniotic fluid-derived mucin. Fetal colonic mucosal cells express the sialyl Tn antigen, particularly in goblet cell mucin. The reactivities of these moABs with a perchloric acid extract of meconium (meconium extract) and different native and neuraminidase treated glycoproteins were examined by solid-phase enzyme-linked immunosorbent assay (ELISA). All the moABs react with the meconium supernatant and meconium extract. The reactivities of TKH-2, MA54, and MA61 are neuraminidase sensitive, and the reactivity of TKH-2 with meconium extract was specifically inhibited by ovine submaxillary mucin (OSM). A NeuAc alpha 2-6GalNAc epitope is the characteristic component in meconium. Mucin released from the fetal respiratory tract could, in part, provide an alternative source in the amniotic fluid. TKH-2 is the most sensitive antibody directed to sialyl Tn antigen in meconium supernatant. The likelihood of TKH-2 serving as the basis for a sensitive assay to detect sialyl Tn in meconium- and amniotic fluid-derived mucin is provided.
Kobayashi
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Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Shizuoka, Japan.
Ohi
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Terao
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D000653:Amniotic Fluid / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000302:isolation & purification; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005333:Fetus / Q000276:immunology; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology; D008470:Meconium / Q000276:immunology*; D009077:Mucins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D009077:Mucins; C055491:sialosyl-Tn antigen