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Recombining germline-derived CDR sequences for creating diverse single-framework antibody libraries

Söderlind, Eskil LU ; Strandberg, Leif LU ; Jirholt, Pernilla LU ; Kobayashi, Norihiro; Alexeiva, Vessela; Åberg, Anna Maria LU ; Nilsson, Anna; Jansson, Bo LU ; Ohlin, Mats LU and Wingren, Christer LU , et al. (2000) In Nature Biotechnology 18(8). p.852-856
Abstract

We constructed a single-chain Fv antibody library that permits human complementarity-determining region (CDR) gene fragments of any germline to be incorporated combinatorially into the appropriate positions of the variable-region frameworks VH-DP47 and VL-DPL3. A library of 2 x 109 independent transformants was screened against haptens, peptides, carbohydrates, and proteins, and the selected antibody fragments exhibited dissociation constants in the subnanomolar range. The antibody genes in this library were built on a single master framework into which diverse CDRs were allowed to recombine. These CDRs were sampled from in vivo-processed gene sequences, thus potentially optimizing the levels of correctly folded and... (More)

We constructed a single-chain Fv antibody library that permits human complementarity-determining region (CDR) gene fragments of any germline to be incorporated combinatorially into the appropriate positions of the variable-region frameworks VH-DP47 and VL-DPL3. A library of 2 x 109 independent transformants was screened against haptens, peptides, carbohydrates, and proteins, and the selected antibody fragments exhibited dissociation constants in the subnanomolar range. The antibody genes in this library were built on a single master framework into which diverse CDRs were allowed to recombine. These CDRs were sampled from in vivo-processed gene sequences, thus potentially optimizing the levels of correctly folded and functional molecules, and resulting in a molecule exhibiting a lower computed immunogenicity compared to naive immunoglobulins. Using the modularized assembly process to incorporate foreign sequences into an immunoglobulin scaffold, it is possible to vary as many as six CDRs at the same time, creating genetic and funcfional variation in antibody molecules.

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organization
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type
Contribution to journal
publication status
published
keywords
Antibody engineering, Diversity, In vitro evolution, Recombination
in
Nature Biotechnology
volume
18
issue
8
pages
5 pages
publisher
Nature Publishing Group
external identifiers
  • Scopus:0033883409
ISSN
1087-0156
DOI
10.1038/78458
language
English
LU publication?
yes
id
2f40154d-08d6-406d-80e4-e58ddb724997
date added to LUP
2016-04-19 14:03:45
date last changed
2016-12-04 04:50:37
@misc{2f40154d-08d6-406d-80e4-e58ddb724997,
  abstract     = {<p>We constructed a single-chain Fv antibody library that permits human complementarity-determining region (CDR) gene fragments of any germline to be incorporated combinatorially into the appropriate positions of the variable-region frameworks VH-DP47 and VL-DPL3. A library of 2 x 10<sup>9</sup> independent transformants was screened against haptens, peptides, carbohydrates, and proteins, and the selected antibody fragments exhibited dissociation constants in the subnanomolar range. The antibody genes in this library were built on a single master framework into which diverse CDRs were allowed to recombine. These CDRs were sampled from in vivo-processed gene sequences, thus potentially optimizing the levels of correctly folded and functional molecules, and resulting in a molecule exhibiting a lower computed immunogenicity compared to naive immunoglobulins. Using the modularized assembly process to incorporate foreign sequences into an immunoglobulin scaffold, it is possible to vary as many as six CDRs at the same time, creating genetic and funcfional variation in antibody molecules.</p>},
  author       = {Söderlind, Eskil and Strandberg, Leif and Jirholt, Pernilla and Kobayashi, Norihiro and Alexeiva, Vessela and Åberg, Anna Maria and Nilsson, Anna and Jansson, Bo and Ohlin, Mats and Wingren, Christer and Danielsson, Lena and Carlsson, Roland and Borrebaeck, Carl A K},
  issn         = {1087-0156},
  keyword      = {Antibody engineering,Diversity,In vitro evolution,Recombination},
  language     = {eng},
  number       = {8},
  pages        = {852--856},
  publisher    = {ARRAY(0xa7503d0)},
  series       = {Nature Biotechnology},
  title        = {Recombining germline-derived CDR sequences for creating diverse single-framework antibody libraries},
  url          = {http://dx.doi.org/10.1038/78458},
  volume       = {18},
  year         = {2000},
}