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^a Departments of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA

^bOncoImmune, Inc. Columbus, OH 43212, USA

^c Department of Surgery and Internal Medicine, University of Michigan, Ann Harbor, MI 48109, USA

^d Center for Retrovirus Research and Comprehensive Cancer Center, College of Pharmacy, The Ohio State University, Columbus, OH 43210, USA

^e Department of Molecular Virology, Immunology & Medical Genetics, School of Biomedical Science, College of Medicine, The Ohio State University, Columbus, OH 43210, USA

*Corresponding author. Fax: 1-614-688-3106

Email address:  wang.892@osu.edu;  yangl@med.umich.edu;  kvaratskhelia.1@osu.edu;  freitas.5@osu.edu

Abstract

CD24 is a glycosyl-phosphatidyl-inositol linked glycoprotein expressed in a broad range of cell types and is heavily glycosylated. It has been found to be over expressed in cancers and tumors and is also a costimulatory molecule. Therefore, this study was carried out to define the structures of the carbohydrates associated with the CD24 recombinant protein. The CD24 glycoprotein's oligosaccharides were released by chemical and enzymatic means prior to being analyzed by MALDI-TOF-MS. The results obtained showed that CD24 is both N- and O-glycosylated. The major oligosaccharides were found to be Neu5Aca-2,3/6Galβ-1,3GalNAc, NeuAc 2 Gal β-1,3GalNAc 1 (O-glycans) , GalNAc 2 GlcNAc 2 Man 3 Fuc 1, Gal 1 GalNAc 2 GlcNAc 2 Man 3 Fuc 1 , and Gal 2 GalNAc 2 GlcNAc 2 Man 3 Fuc 1 (N-glycans). The results showed that Neu5Aca-2,3/6Galβ-1,3GalNAc (sialyl-tumor antigen, sT), a cancer-associated carbohydrate, was the most abundant glycan associated with CD24. This result raised the intriguing possibility that CD24 may be a major carrier of the sialyl-T abundantly found in cancer cells.

Keywords: CD24; Glycans; MALDI-TOF-MS; T antigen

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