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A focused antibody library for improved hapten recognition

Persson, Helena LU ; Lantto, Johan LU and Ohlin, Mats LU orcid (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:
author
; and
organization
publishing date
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
2021-06-08 06:01:43
@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},
  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},
}