Skip Navigation



Glycobiology Advance Access published online on May 4, 2005
Glycobiology, doi:10.1093/glycob/cwi072
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrow All Versions of this Article:
15/9/849    most recent
cwi072v1
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 Liners, F.
Right arrow Articles by Van Cutsem, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Liners, F.
Right arrow Articles by Van Cutsem, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oupjournals.org
Received January 12, 2005
Revised April 6, 2005
Accepted April 30, 2005

Article

Production and characterization of a phage-display recombinant antibody against carrageenans. Evidence for the recognition of a secondary structure of carrageenan chains present in red algae tissues

Françoise Liners 1, William Helbert 2, and Pierre Van Cutsem 1*

1 URBV, University of Namur, Rue de Bruxelles 61, 5000 Namur, Belgium
2 UMR7139 CNRSGoëmar-UPMC « Végétaux Marins et Biomolécules », Station Biologique de Roscoff, Pl. G.Teissier29, 680 Roscoff, France

* To whom correspondence should be addressed.
Pierre Van Cutsem, E-mail: pierre.vancutsem{at}fundp.ac.be


  Abstract

We report the isolation, for the first time by phage display, of a scFv recombinant antibody called B3 directed against carrageenans, the major sulphated polysaccharides of red seaweeds. Immunoassays were used to characterize the binding of B3 antibodies towards the three main carrageenan forms ({kappa}, {iota} and {lambda}) differing by their sulfonic ester content and the presence of 3,6-anhydrogalactose. In ELISA, B3 soluble scFv showed a high reactivity towards {iota}-carrageenan at any titer but, at high titer only, recognized also the highly sulfated {lambda}-form. Surface-adsorbed {kappa}-polymers were only recognized in presence of poly-L-lysine. The replacement of Na+ ions by K+ in the buffers had no effect on {kappa}-polymer detection but increased the binding of B3 antibodies towards both {iota}-and {lambda}-carrageenans, whereas addition of Ca2+ decreased sharply the recognition of the {iota}-form. In competitive assays, low titer B3 soluble scFv showed a {iota}>{kappa}>{lambda} selectivity and recognized a mixture of {iota}-oligomers with degrees of polymerization between 4 and 18 but not sub-fractions of 4 or 6 residues long. We suggest therefore that the B3 epitope could consist of a helical conformation of carrageenan chains. Immunofluorescence microscopy showed that, amongst other red algae, Chondrus gametophyte (containing {iota}-chains) was strongly recognized by B3 scFv whereas sporophytic tissues rich in {lambda}-carrageenans were not, assessing the preference of this probe for {iota}-carrageenans in situ. The high potential of the B3 recombinant probe is discussed.

Keywords: carrageenan/ helix/ immunolabeling/ phage display/ red algae.
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.