title: A Cytotoxic Antibody Recognizing Lacto-N-fucopentaose I (LNFP I) on Human Induced Pluripotent Stem (hiPS) Cells.
abstract: We have generated a mouse monoclonal antibody (R-17F, IgG1 subtype) specific to human induced pluripotent stem (hiPS)/embryonic stem (ES) cells by using a hiPS cell line as an antigen. Triple-color confocal immunostaining images of hiPS cells with R-17F indicated that the R-17F epitope was expressed exclusively and intensively on the cell membranes of hiPS cells and co-localized partially with those of SSEA-4 and SSEA-3. Lines of evidence suggested that the predominant part of the R-17F epitope was a glycolipid. Upon TLC blot of total lipid extracts from hiPS cells with R-17F, one major R-17F-positive band was observed at a slow migration position close to that of anti-blood group H1(O) antigen. MALDI-TOF-MS and MS(n) analyses of the purified antigen indicated that the presumptive structure of the R-17F antigen was Fuc-Hex-HexNAc-Hex-Hex-Cer. Glycan microarray analysis involving 13 different synthetic oligosaccharides indicated that R-17F bound selectively to LNFP I (Fucα1-2Galβ1-3GlcNAcβ1-3Galβ1-4Glc). A critical role of the terminal Fucα1-2 residue was confirmed by the selective disappearance of R-17F binding to the purified antigen upon α1-2 fucosidase digestion. Most interestingly, R-17F, when added to hiPS/ES cell suspensions, exhibited potent dose-dependent cytotoxicity. The cytotoxic effect was augmented markedly upon the addition of the secondary antibody (goat anti-mouse IgG1 antibody). R-17F may be beneficial for safer regenerative medicine by eliminating residual undifferentiated hiPS cells in hiPS-derived regenerative tissues, which are considered to be a strong risk factor for carcinogenesis.
authors.affiliation: From the Research Center for Glycobiotechnology, Ritsumeikan University, Noji-Higashi, 1-1-1, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Matsumoto
authors.affiliation: From the Research Center for Glycobiotechnology, Ritsumeikan University, Noji-Higashi, 1-1-1, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Nakao
authors.affiliation: From the Research Center for Glycobiotechnology, Ritsumeikan University, Noji-Higashi, 1-1-1, Kusatsu, Shiga 525-8577, Japan, the Laboratory of Stem Cell Cultures and.
authors.forename: Keiko
authors.initials: K
authors.lastname: Kawabe
authors.affiliation: From the Research Center for Glycobiotechnology, Ritsumeikan University, Noji-Higashi, 1-1-1, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Motohiro
authors.initials: M
authors.lastname: Nonaka
authors.affiliation: the Laboratory of Bio-analytical Chemistry, College of Pharmaceutical Sciences, Ritsumeikan University, Shiga 525-8577, Japan.
authors.forename: Hidenao
authors.initials: H
authors.lastname: Toyoda
authors.affiliation: the Laboratory of Bio-analytical Chemistry, College of Pharmaceutical Sciences, Ritsumeikan University, Shiga 525-8577, Japan.
authors.forename: Yuto
authors.initials: Y
authors.lastname: Takishima
authors.affiliation: the Laboratory of Stem Cell Regulation, National Institutes of Biomedical Innovation, Health, and Nutrition, Osaka 567-0085, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kawabata
authors.affiliation: the Laboratory of Stem Cell Regulation, National Institutes of Biomedical Innovation, Health, and Nutrition, Osaka 567-0085, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamaguchi
authors.affiliation: the Laboratory of Stem Cell Cultures and.
authors.forename: Miho K
authors.initials: MK
authors.lastname: Furue
authors.affiliation: the Niigata University of Pharmacy and Applied Life Sciences, Niigata 956-0841, Japan, AGT&T Co. Ltd., Tokushima 771-1151, Japan, and.
authors.forename: Takao
authors.initials: T
authors.lastname: Taki
authors.affiliation: the Analytical and Measuring Instruments Division, Shimadzu Corp., Kyoto 604-8511, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Okumura
authors.affiliation: the Analytical and Measuring Instruments Division, Shimadzu Corp., Kyoto 604-8511, Japan.
authors.forename: Yuzo
authors.initials: Y
authors.lastname: Yamazaki
authors.affiliation: the Analytical and Measuring Instruments Division, Shimadzu Corp., Kyoto 604-8511, Japan.
authors.forename: Shuuichi
authors.initials: S
authors.lastname: Nakaya
authors.affiliation: From the Research Center for Glycobiotechnology, Ritsumeikan University, Noji-Higashi, 1-1-1, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Nobuko
authors.initials: N
authors.lastname: Kawasaki
authors.affiliation: From the Research Center for Glycobiotechnology, Ritsumeikan University, Noji-Higashi, 1-1-1, Kusatsu, Shiga 525-8577, Japan, tkawasak@fc.ritsumei.ac.jp.
authors.forename: Toshisuke
authors.initials: T
authors.lastname: Kawasaki
mesh_terms: D000906:Antibodies / Q000276:immunology*; D002240:Carbohydrate Sequence; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000276:immunology*
chemical_list: D000906:Antibodies; D009844:Oligosaccharides; C045442:lacto-N-fucopentaose I
keywords: MALDI-TOF-MS; carbohydrate-recognizing antibody; confocal microscopy; cytotoxic antibody; embryonic stem cell; flow cytometry; glycan microarray; glycolipid; induced pluripotent stem cell (iPS cell) (iPSC); lacto-N-fucopentaose I