Skip Navigation


Glycobiology Advance Access originally published online on March 1, 2007
Glycobiology 2007 17(5):529-540; doi:10.1093/glycob/cwm017
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrowOA All Versions of this Article:
17/5/529    most recent
cwm017v2
cwm017v1
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 ISI Web of Science
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 (9)
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Jones, A. J.S.
Right arrow Articles by Battersby, J. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jones, A. J.S.
Right arrow Articles by Battersby, J. E.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Selective clearance of glycoforms of a complex glycoprotein pharmaceutical caused by terminal N-acetylglucosamine is similar in humans and cynomolgus monkeys

Andrew J.S. Jones4, Damon I. Papac2,4, Edward H. Chin4, Rodney Keck4, Sharon A. Baughman3,5, Yvonne S. Lin5, Johannes Kneer6 and John E. Battersby1,4

4 Department of Analytical Chemistry
5 Department of Pharmacokinetics and Metabolism, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080
6 Department of Clinical Pharmacology, F. Hoffmann–La Roche Ltd, CH-4070, Basel, Switzerland

1 To whom correspondence should be addressed: Tel: 650 225 6264; Fax: 650 225 3554; e-mail: jeb{at}gene.com

Received on November 17, 2006; revised on February 10, 2007; accepted on February 11, 2007

To understand how the carbohydrate moieties of a recombinant glycoprotein affected its pharmacokinetic (PK) properties, the glycan distribution was directly assessed from serial blood samples taken during PK studies in cynomolgus monkeys and humans. The protein studied was an immunoadhesin (lenercept), containing an Fc domain from human immunoglobulin G (IgG-1) and two copies of the extensively glycosylated extra cellular domain of tumor necrosis factor receptor p55. The protein was recovered in pure form using a dual column, immunoaffinity-reversed-phase high-performance liquid chromatography method. The glycans were released and analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). Alternatively, trypsin was used to obtain glycopeptides, and these were analyzed by MALDI-TOF. The composition versus time profiles show that the distribution of glycans in the Fc domain was not altered over 10 days of circulation, consistent with their sequestration in the interior of the protein. However, the glycan composition in the receptor domain was changed dramatically in the first 24 h and then remained relatively constant. Analysis of the acidic glycans (derived exclusively from the receptor domain) showed that, in the rapid initial phase of clearance, glycans carrying terminal N-acetylglucosamine (tGlcNAc) were selectively cleared from the circulation. This phenomenon occurred similarly in humans and cynomolgus monkeys. Sialic acid content and terminal galactose showed only small changes. These data confirm the correlation of tGlcNAc and half-life of the molecule, and support the hypothesis that the mannose receptor (which can also bind tGlcNAc) causes the variable clearance of this molecule.

Key words: immunoadhesin / lenercept / terminal N-acetylglucosamine / selective glycan clearance / IgG1 Fc glycans / terminal galactose

2 Present address: Myriad Pharmacueticals, Inc., Salt Lake City, UT 84108

3 Present address: Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320-1799


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
X. Chen, Y D. Liu, and G. C Flynn
The effect of Fc glycan forms on human IgG2 antibody clearance in humans
Glycobiology, March 1, 2009; 19(3): 240 - 249.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
G.-C. Gil, W. H Velander, and K. E Van Cott
Analysis of the N-glycans of recombinant human Factor IX purified from transgenic pig milk
Glycobiology, July 1, 2008; 18(7): 526 - 539.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. S. Powlesland, T. Fisch, M. E. Taylor, D. F. Smith, B. Tissot, A. Dell, S. Pohlmann, and K. Drickamer
A Novel Mechanism for LSECtin Binding to Ebola Virus Surface Glycoprotein through Truncated Glycans
J. Biol. Chem., January 4, 2008; 283(1): 593 - 602.
[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.