Skip Navigation


Glycobiology Advance Access originally published online on July 16, 2008
Glycobiology 2008 18(10):789-798; doi:10.1093/glycob/cwn068
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
18/10/789    most recent
cwn068v3
cwn068v2
cwn068v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (4)
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Tateno, H.
Right arrow Articles by Hirabayashi, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tateno, H.
Right arrow Articles by Hirabayashi, J.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Glycoconjugate microarray based on an evanescent-field fluorescence-assisted detection principle for investigation of glycan-binding proteins

Hiroaki Tateno, Atsushi Mori, Noboru Uchiyama, Rikio Yabe, Jun Iwaki, Toshihide Shikanai, Takashi Angata, Hisashi Narimatsu and Jun Hirabayashi1

Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Central 2, 1-1-1 Umezono, Ibaraki 305-8568, Japan

1 To whom correspondence should be addressed: Tel: +81-29-861-3124; Fax: +81-29-861-3125; e-mail: jun-hirabayashi{at}aist.go.jp

Received on April 29, 2008; revised on June 10, 2008; accepted on July 8, 2008

The extensive involvement of glycan-binding proteins (GBPs) as regulators in diverse biological phenomena provides a fundamental reason to investigate their glycan-binding specificities. Here, we developed a glycoconjugate microarray based on an evanescent-field fluorescence-assisted detection principle for investigation of GBPs. Eighty-nine selected multivalent glycoconjugates comprising natural glycoproteins, neo-glycoproteins, and polyacrylamide (PAA)-conjugated glycan epitopes were immobilized on an epoxy-activated glass slide. The GBP binding was monitored by an evanescent-field fluorescence-assisted scanner at equilibrium without washing steps. The detection principle also allows direct application of unpurified GBPs with the aid of specific antibodies. Model experiments using plant lectins (RCA120, ConA, and SNA), galectins (3 and 8), a C-type lectin (DC-SIGN) and a siglec (CD22) provided data consistent with previous work within 4 h using less than 40 ng of GBPs per analysis. As an application, serum profiling of antiglycan antibodies (IgG and IgM) was performed with Cy3-labeled secondary antibodies. Moreover, novel carbohydrate-binding ability was demonstrated for a human IL-18 binding protein. Thus, the developed glycan array is useful for investigation of various types of GBPs, with the added advantage of wash-free analysis.

Key words: evanescent / glycoconjugate / lectin / microarray / specificity


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 GlycobiologyHome page
A. Marson, D. E Robinson, P. N Brookes, B. Mulloy, M. Wiles, S. J Clark, H. L Fielder, L. J Collinson, S. A Cain, C. M Kielty, et al.
Development of a microtiter plate-based glycosaminoglycan array for the investigation of glycosaminoglycan-protein interactions
Glycobiology, December 1, 2009; 19(12): 1537 - 1546.
[Abstract] [Full Text] [PDF]

Home page
 J BiochemHome page
K. Mitsunaga, J. Harada-Itadani, T. Shikanai, H. Tateno, Y. Ikehara, J. Hirabayashi, H. Narimatsu, and T. Angata
Human C21orf63 is a Heparin-binding Protein
J. Biochem., September 1, 2009; 146(3): 369 - 373.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Yamasaki, M. Matsumoto, O. Takeuchi, T. Matsuzawa, E. Ishikawa, M. Sakuma, H. Tateno, J. Uno, J. Hirabayashi, Y. Mikami, et al.
C-type lectin Mincle is an activating receptor for pathogenic fungus, Malassezia
PNAS, February 10, 2009; 106(6): 1897 - 1902.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.