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Mucin, a major component of mucus, is a highly O-glycosylated, high-molecular-mass glycoprotein extensively involved in the physiology of gastrointestinal mucosa. To detect and characterize mucins derived from site-specific mucous cells, we developed a monoclonal antibody, designated PGM34, by immunizing a mouse with purified pig gastric mucin. | ỊFEBS Journal A monoclonal antibody PGM34 against 6-sulfated blood-group H type 2 antigen on the carbohydrate moiety of mucin Analysis of the epitope sequence and immunohistochemical study Daigo Tsubokawa1 Yukinobu Goso1 Akira Sawaguchi2 Makoto Kurihara3 Takafumi Ichikawa4 Noriko Sato5 Tatsuo Suganuma2 Kyoko Hotta4 and Kazuhiko Ishihara1 1 Department of Biochemistry Kitasato University Graduate Schoolof MedicalSciences Sagamihara Japan 2 Department of Anatomy UltrastructuralCellBiology Faculty of Medicine University of Miyazaki Japan 3 Isehara Research Laboratory Kanto ChemicalCo. Inc. Isehara Japan 4 Department of Biochemistry Kitasato University Schoolof Medicine Sagamihara Japan 5 Department of InstrumentalAnalysis Kitasato University Schoolof PharmaceuticalSciences Tokyo Japan Keywords monoclonalantibody mucin mucous cells sulfated oligosaccharide Correspondence K. Ishihara Department of Biochemistry Kitasato University Schoolof Allied Health Sciences 1-15-1 Kitasato Sagamihara 2288555 Japan Fax Tel 81 42 778 8262 E-mail isiharak@kitasato-u.ac.jp Received 9 November 2006 revised 16 January 2007 accepted 7 February 2007 doi 10.1111 j.1742-4658.2007.05731.x Mucin a major component of mucus is a highly O-glycosylated high-molecular-mass glycoprotein extensively involved in the physiology of gastrointestinal mucosa. To detect and characterize mucins derived from site-specific mucous cells we developed a monoclonal antibody designated PGM34 by immunizing a mouse with purified pig gastric mucin. The reactivity of PGM34 with mucin was inhibited by periodate treatment of the mucin but not by trypsin digestion. This suggests that PGM34 recognizes the carbohydrate portion of mucin. To determine the epitope oligosaccha-ride-alditols obtained from pig gastric mucin were fractionated by successive gel-filtration ion-exchange and normal-phase HPLC and tested for reactivity with PGM34. Two purified oligosaccharide-alditols that reacted with PGM34 were obtained. Their .
