Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
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 arrowRequest Permissions
Google Scholar
Right arrow Articles by Hugo, N.
Right arrow Articles by Altschuh, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hugo, N.
Right arrow Articles by Altschuh, D.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Protein Engineering vol. 16 no. 5 pp. 381-386, 2003
© 2003 Oxford University Press

VL position 34 is a key determinant for the engineering of stable antibodies with fast dissociation rates

N. Hugo1, M. Weidenhaupt2, M. Beukes1, B. Xu3, J.-C. Janson3, T. Vernet2 and D. Altschuh1,4

1UMR 7100, ‘Biotechnologie des Interactions Macromoléculaires’, Ecole Supérieure de Biotechnologie de Strasbourg (ESBS), Parc d’Innovation, boulevard Sébastien Brant, BP 10413, F-67412 Illkirch Cedex, 2Laboratoire d’Ingénierie des Macromolécules, Institut de Biologie Structurale Jean-Pierre Ebel (CEA/CNRS/UJF), 41 avenue Jules Horowitz, F-38027 Grenoble Cedex 1, France and 3Center for Surface Biotechnology, Uppsala Biomedical Center, Box 577, S-751 23 Uppsala, Sweden N.Hugo and M.Weidenhaupt contributed equally to this work.

4 To whom correspondence should be addressed. e-mail: daniele.altschuh{at}esbs.u-strasbg.fr

Predictive engineering of antibodies exhibiting fast kinetic properties could provide reagents for biotechnological applications such as continuous monitoring of compounds or affinity chromatography. Based on covariance analysis of murine germline antibody variable domains, we selected position L34 (Kabat numbering) for mutational studies. This position is located at the VL/VH interface, at the base of the paratope but with limited antigen contacts, thus making it an attractive position for mild alterations of antigen binding properties. We introduced a serine at position L34 in two different antibodies: Fab (fragment antigen binding) 57P (Asn34Ser) and scFv (single chain fragment variable) 1F4 (Gln34Ser), that recognize peptides derived from the coat protein of tobacco mosaic virus and the oncoprotein E6, respectively. Both mutated antibodies exhibited similar properties: (i) expression levels of active fragments in Escherichia coli were markedly improved; (ii) thermostability was enhanced; and (iii) dissociation rate parameters (koff) were increased by 2- and at least 57-fold for scFv1F4 and Fab57P, respectively, while their association rate parameters (kon) remained unchanged. The L34 Ala and Thr mutants of both antibody fragments did not possess these properties. This first demontration of similar effects observed in two antibodies with different specificities may open the way to the predictive design of molecules with enhanced stability and fast dissociation rates.

Received October 3, 2002; revised March 7, 2003; accepted April 4, 2003.


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.