A focused antibody library for improved hapten recognition
(2006) In Journal of Molecular Biology 357(2). p.607-620- Abstract
- The topography of the antigen-binding site as well as the number and the positioning of the antigen contact residues are strongly correlated with the size of the antigen with which the antibody interacts. On the basis of these considerations, we have designed a focused scFv repertoire biased for haptens, designated the cavity library. The hapten-specific scFv, FITC8, was used as a scaffold for library construction. FITC8, like other hapten binders, displays a characteristic cavity in its paratope into which the hapten binds. In five of the six complementarity-determining regions, diversity-carrying residues were selected rationally on the basis of a model structure of FITC8 and on known antibody structure-function relationships, resulting... (More)
- The topography of the antigen-binding site as well as the number and the positioning of the antigen contact residues are strongly correlated with the size of the antigen with which the antibody interacts. On the basis of these considerations, we have designed a focused scFv repertoire biased for haptens, designated the cavity library. The hapten-specific scFv, FITC8, was used as a scaffold for library construction. FITC8, like other hapten binders, displays a characteristic cavity in its paratope into which the hapten binds. In five of the six complementarity-determining regions, diversity-carrying residues were selected rationally on the basis of a model structure of FITC8 and on known antibody structure-function relationships, resulting in variation of 11 centrally located, cavity-lining residues. L3 was allowed to carry a more complex type of diversity. In addition, length variation was introduced into H2, as longer versions of this loop have been shown to correlate with increased hapten binding. The library was screened, using phage display, against a panel of five different haptens, yielding diverse and highly specific binders to four of the antigens. Parallel selections were performed with a library having diversity spread onto a greater area, including more peripherally located residues. This resulted in the isolation of binders, which, in contrast to the clones selected from the cavity library, were not able to bind to the soluble hapten in the absence of the carrier protein. Thus, we have shown that by focusing diversity to the hotspots of interaction a library with improved hapten-binding ability can be created. The study supports the notion that it is possible to create antibody libraries that are biased for the recognition of antigens of pre-defined size. (c) 2006 Elsevier Ltd. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/415689
- author
- Persson, Helena LU ; Lantto, Johan LU and Ohlin, Mats LU
- organization
- publishing date
- 2006
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- display, phage, hapten, focused diversity, antibody evolution, antibody library
- in
- Journal of Molecular Biology
- volume
- 357
- issue
- 2
- pages
- 607 - 620
- publisher
- Elsevier
- external identifiers
-
- wos:000236120200022
- pmid:16445941
- scopus:33344476078
- pmid:16445941
- ISSN
- 1089-8638
- DOI
- 10.1016/j.jmb.2006.01.004
- language
- English
- LU publication?
- yes
- id
- 1331a949-5122-410b-867b-7ff2890e7f92 (old id 415689)
- date added to LUP
- 2016-04-01 15:44:25
- date last changed
- 2022-01-28 06:49:29
@article{1331a949-5122-410b-867b-7ff2890e7f92, abstract = {{The topography of the antigen-binding site as well as the number and the positioning of the antigen contact residues are strongly correlated with the size of the antigen with which the antibody interacts. On the basis of these considerations, we have designed a focused scFv repertoire biased for haptens, designated the cavity library. The hapten-specific scFv, FITC8, was used as a scaffold for library construction. FITC8, like other hapten binders, displays a characteristic cavity in its paratope into which the hapten binds. In five of the six complementarity-determining regions, diversity-carrying residues were selected rationally on the basis of a model structure of FITC8 and on known antibody structure-function relationships, resulting in variation of 11 centrally located, cavity-lining residues. L3 was allowed to carry a more complex type of diversity. In addition, length variation was introduced into H2, as longer versions of this loop have been shown to correlate with increased hapten binding. The library was screened, using phage display, against a panel of five different haptens, yielding diverse and highly specific binders to four of the antigens. Parallel selections were performed with a library having diversity spread onto a greater area, including more peripherally located residues. This resulted in the isolation of binders, which, in contrast to the clones selected from the cavity library, were not able to bind to the soluble hapten in the absence of the carrier protein. Thus, we have shown that by focusing diversity to the hotspots of interaction a library with improved hapten-binding ability can be created. The study supports the notion that it is possible to create antibody libraries that are biased for the recognition of antigens of pre-defined size. (c) 2006 Elsevier Ltd. All rights reserved.}}, author = {{Persson, Helena and Lantto, Johan and Ohlin, Mats}}, issn = {{1089-8638}}, keywords = {{display; phage; hapten; focused diversity; antibody evolution; antibody library}}, language = {{eng}}, number = {{2}}, pages = {{607--620}}, publisher = {{Elsevier}}, series = {{Journal of Molecular Biology}}, title = {{A focused antibody library for improved hapten recognition}}, url = {{http://dx.doi.org/10.1016/j.jmb.2006.01.004}}, doi = {{10.1016/j.jmb.2006.01.004}}, volume = {{357}}, year = {{2006}}, }