Next Article in Journal
Identification and Biochemical Characterization of High Mobility Group Protein 20A as a Novel Ca2+/S100A6 Target
Previous Article in Journal
Effects on Lignin Redistribution in Eucalyptus globulus Fibres Pre-Treated by Steam Explosion: A Microscale Study to Cellulose Accessibility
Previous Article in Special Issue
Current Views on the Roles of O-Glycosylation in Controlling Notch-Ligand Interactions
Open AccessArticle

Glycan Epitopes on 201B7 Human-Induced Pluripotent Stem Cells Using R-10G and R-17F Marker Antibodies

1
Laboratory of Bio-analytical Chemistry, College of Pharmaceutical Sciences, Ritsumeikan University, Shiga 525-8577, Japan
2
Glycobiotechnology Laboratory, Ritsumeikan University, Shiga 525-8577, Japan
3
Department of Medical Life Science, Yokohama City University, Kanagawa 230-0045, Japan
*
Author to whom correspondence should be addressed.
Academic Editor: Jun-ichi Furukawa
Biomolecules 2021, 11(4), 508; https://doi.org/10.3390/biom11040508
Received: 7 March 2021 / Accepted: 23 March 2021 / Published: 29 March 2021
(This article belongs to the Special Issue Structural and Functional Approach to the Glycan Diversity)
We developed two human-induced pluripotent stem cell (hiPSC)/human embryonic stem cell (hESC)-specific glycan-recognizing mouse antibodies, R-10G and R-17F, using the Tic (JCRB1331) hiPSC line as an antigen. R-10G recognizes a low-sulfate keratan sulfate, and R-17F recognizes lacto-N-fucopentaose-1. To evaluate the general characteristics of stem cell glycans, we investigated the hiPSC line 201B7 (HPS0063), a prototype iPSC line. Using an R-10G affinity column, an R-10G-binding protein was isolated from 201B7 cells. The protein yielded a single but very broad band from 480 to 1236 kDa by blue native gel electrophoresis. After trypsin digestion, the protein was identified as podocalyxin by liquid chromatography/mass spectrometry. According to Western blotting, the protein reacted with R-10G and R-17F. The R-10G-positive band was resistant to digestion with glycan-degrading enzymes, including peptide N-glycanase, but the intensity of the band was decreased significantly by digestion with keratanase, keratanase II, and endo-β-galactosidase, suggesting the R-10G epitope to be a keratan sulfate. These results suggest that keratan sulfate-type epitopes are shared by hiPSCs. However, the keratan sulfate from 201B7 cells contained a polylactosamine disaccharide unit (Galβ1-4GlcNAc) at a significant frequency, whereas that from Tic cells consisted mostly of keratan sulfate disaccharide units (Galβ1-4GlcNAc(6S)). In addition, the abundance of the R-10G epitope was significantly lower in 201B7 cells than in Tic cells. View Full-Text
Keywords: human-induced pluripotent stem cells (hiPSCs); monoclonal antibodies; R-10G; R-17F; keratan sulfate; podocalyxin; keratanase II; endo-β-galactosidase human-induced pluripotent stem cells (hiPSCs); monoclonal antibodies; R-10G; R-17F; keratan sulfate; podocalyxin; keratanase II; endo-β-galactosidase
Show Figures

Figure 1

MDPI and ACS Style

Nagai, Y.; Nakao, H.; Kojima, A.; Komatsubara, Y.; Ohta, Y.; Kawasaki, N.; Kawasaki, N.; Toyoda, H.; Kawasaki, T. Glycan Epitopes on 201B7 Human-Induced Pluripotent Stem Cells Using R-10G and R-17F Marker Antibodies. Biomolecules 2021, 11, 508. https://doi.org/10.3390/biom11040508

AMA Style

Nagai Y, Nakao H, Kojima A, Komatsubara Y, Ohta Y, Kawasaki N, Kawasaki N, Toyoda H, Kawasaki T. Glycan Epitopes on 201B7 Human-Induced Pluripotent Stem Cells Using R-10G and R-17F Marker Antibodies. Biomolecules. 2021; 11(4):508. https://doi.org/10.3390/biom11040508

Chicago/Turabian Style

Nagai, Yuko; Nakao, Hiromi; Kojima, Aya; Komatsubara, Yuka; Ohta, Yuki; Kawasaki, Nana; Kawasaki, Nobuko; Toyoda, Hidenao; Kawasaki, Toshisuke. 2021. "Glycan Epitopes on 201B7 Human-Induced Pluripotent Stem Cells Using R-10G and R-17F Marker Antibodies" Biomolecules 11, no. 4: 508. https://doi.org/10.3390/biom11040508

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop