Skip Navigation



Glycobiology Advance Access published online on February 23, 2006
Glycobiology, doi:10.1093/glycob/cwj091
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrow All Versions of this Article:
16/6/514    most recent
cwj091v1
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 arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Wang, Y.
Right arrow Articles by Hancock, W. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wang, Y.
Right arrow Articles by Hancock, W. S.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
Received November 13, 2005
Revised January 23, 2006
Accepted February 13, 2006

Article

Approaches to the Study of N-linked Glycoproteins in Human Plasma Using Lectin Affinity Chromatography and Nano-HPLC Coupled to Electrospray Linear Ion Trap - Fourier Transform Mass Spectrometry (LTQ/FTMS)

Yonghui Wang 1, Shiaw-lin Wu 1, and William S. Hancock 1 *

1 Barnett Institute, Northeastern University, Boston, MA 02115

* To whom correspondence should be addressed.
William S. Hancock, E-mail: wi.hancock{at}neu.edu


  Abstract

In this publication, we will describe the combination of lectin affinity chromatography with nano HPLC coupled to a linear ion trap Fourier transform mass spectrometer (capillary LC-LTQ/FTMS) to characterize N-linked glycosylation structures in human plasma proteins. We used a well characterized glycoprotein, tissue plasminogen activator (rt-PA), which is present at low levels in blood, as a standard to determine the dynamic range of this approach. N-linked glycopeptides derived from rt-PA could be characterized at a ratio of 1:200 in human plasma (rtPA:total plasma protein, w/w) by accurate mass measurement in the FTMS and fragmentation (MSn) in the linear ion trap. We demonstrated that this platform has the potential to characterize the general N-linked glycosylation structures of abundant glycoproteins present in human plasma without the requirement for antibody based purification or additional carbohydrate analytical protocols. This conclusion was supported by the determination of carbohydrate structures for three glycoproteins, IgG, haptoglobin and alpha-1-acid glycoprotein, at their natural levels in a human plasma sample, but only after the lectin enrichment step.

Keywords: lectin/glycoprotein/mass spectrometry/plasma/Chinese hamster ovary.
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
 Eukaryot CellHome page
D. M. Ratner, J. Cui, M. Steffen, L. L. Moore, P. W. Robbins, and J. Samuelson
Changes in the N-Glycome, Glycoproteins with Asn-Linked Glycans, of Giardia lamblia with Differentiation from Trophozoites to Cysts
Eukaryot. Cell, November 1, 2008; 7(11): 1930 - 1940.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
F. Fenaille, M. Le Mignon, C. Groseil, C. Ramon, S. Riande, L. Siret, and N. Bihoreau
Site-specific N-glycan characterization of human complement factor H
Glycobiology, September 1, 2007; 17(9): 932 - 944.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
R. R. Drake, E. E. Schwegler, G. Malik, J. Diaz, T. Block, A. Mehta, and O. J. Semmes
Lectin Capture Strategies Combined with Mass Spectrometry for the Discovery of Serum Glycoprotein Biomarkers
Mol. Cell. Proteomics, October 1, 2006; 5(10): 1957 - 1967.
[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.