Skip Navigation



Glycobiology Advance Access published online on September 25, 2006
Glycobiology, doi:10.1093/glycob/cwl053
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrow All Versions of this Article:
17/1/36    most recent
cwl053v1
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 Kogelberg, H.
Right arrow Articles by Chester, K. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kogelberg, H.
Right arrow Articles by Chester, K. A.
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 June 22, 2006
Revised August 29, 2006
Accepted September 18, 2006

Article

Clearance Mechanism of a Mannosylated Antibody-Enzyme Fusion Protein used in Experimental Cancer Therapy

Heide Kogelberg 1, Berend Tolner 1, Surinder K. Sharma 1, Mark W. Lowdell 2, Uzma Qureshi 1, Mathew Robson 1, Tim Hillyer 1, R. Barbara Pedley 1, Wouter Vervecken 3, Roland Contreras 3, Richard H.J. Begent 1, and Kerry A. Chester 1 *

1 Cancer Research UK Targeting and Imaging Group, Department of Oncology, Royal Free & University College Medical School, Hampstead Campus, London, NW3 2PF, UK
2 Department of Haematology, Royal Free & University College Medical School, Hampstead Campus, London, NW3 2PF, UK
3 Department of Molecular Biomedical Research, Fundamental and Applied Molecular Biology, Ghent University and Flanders Interuniversity Institute for Biotechnology, Technologiepark 927, B-9052 Ghent-Zwijnaarde, Belgium

* To whom correspondence should be addressed.
Kerry A. Chester, E-mail: k.chester{at}ucl.ac.uk


  Abstract

MFECP1 is a mannosylated antibody-enzyme fusion protein used in antibody-directed enzyme prodrug therapy (ADEPT). The antibody selectively targets tumour cells and the targeted enzyme converts a prodrug into a toxic drug. MFECP1 is obtained from expression in the yeast Pichia pastoris and produced to clinical grade. The P. pastoris derived mannosylation of the fusion protein aids rapid normal tissue clearance required for successful ADEPT. The work presented provides evidence that MFECP1 is cleared by the endocytic and phagocytic mannose receptor (MR), which is known to bind to mannose-terminating glycans. MR transfected fibroblast cells internalize MFECP1 as revealed by flow cytometry and confocal microscopy. Immunofluorescence microscopy shows that in vivo clearance in mice occurs predominantly by MR on liver sinusoidal endothelial cells, although MR is also expressed on adjacent Kupffer cells. In the spleen MFECP1 is taken up by MR-expressing macrophages residing in the red pulp and not by dendritic cells which are found in the marginal zone and white pulp. Clearance can be inhibited in vivo by the MR inhibitor mannan as shown by increased enzyme activities in blood. The work improves understanding of interactions of MFECP1 with normal tissue, shows that glycosylation can be exploited in the design of recombinant anti-cancer therapeutics and opens the ways for optimising pharmacokinetics of mannosylated recombinant therapeutics.

Keywords: Antibody-directed enzyme prodrug therapy (ADEPT); anti-carcinoembryonic antigen (CEA) antibody MFE; clearance; mannose receptor (MR); Mannosylated antibody-enzyme fusion protein MFECP1.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




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.