Skip Navigation


PEDS Advance Access originally published online on May 13, 2008
Protein Engineering Design and Selection 2008 21(8):485-493; doi:10.1093/protein/gzn024
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
21/8/485    most recent
gzn024v1
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 arrowRequest Permissions
Google Scholar
Right arrow Articles by Persson, H.
Right arrow Articles by Ohlin, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Persson, H.
Right arrow Articles by Ohlin, M.
Related Collections
Right arrow 2008
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

In vitro evolution of an antibody fragment population to find high-affinity hapten binders

Helena Persson1, Henrik Wallmark1, Anne Ljungars2, Johan Hallborn2 and Mats Ohlin1,3

1Department of Immunotechnology, Lund University, BMC D13 SE-221 84, Lund 2BioInvent International AB, Sölvegatan 41 SE-223 70, Lund, Sweden

3 To whom correspondence should be addressed. E-mail: mats.ohlin{at}immun.lth.se

Recently, we constructed a focused antibody library tailored to interact with haptens. High functionality of this library was demonstrated, as specific binders could be retrieved to a range of different haptens. In the current study we have developed a mutagenesis and selection strategy in order to further fine-tune the hapten binding properties of these antibody fragments. Testosterone was chosen as model antigen for the investigation. A population, rather than a single clone, originating from this focused library and enriched for testosterone binders, was subjected to random mutagenesis and different phage display selection strategies of various stringencies. These included consecutively lowering the antigen concentration and having, or not having, soluble hapten present during the phage capture and elution steps. The different selection procedures resulted in a considerable increase in apparent affinities for several of the selected populations, from which the highest affinity antibody isolated had a KD of 2 nM, corresponding to an ~200-fold affinity improvement compared with the best clone of the starting population. Importantly, the polyclonal nature of the starting material allowed for the identification of novel unrelated variants that differed in fine-specificity, demonstrating that this approach is valuable for exploring different parts of structure space.

Keywords: affinity maturation/antibody library/hapten/phage display/testosterone

Received April 1, 2008; revised April 1, 2008; accepted April 10, 2008.


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.