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

Author:
  • Eskil Söderlind
  • Leif Strandberg
  • Pernilla Jirholt
  • Norihiro Kobayashi
  • Vessela Alexeiva
  • Anna Maria Åberg
  • Anna Nilsson
  • Bo Jansson
  • Mats Ohlin
  • Christer Wingren
  • Lena Danielsson
  • Roland Carlsson
  • Carl A K Borrebaeck
Publishing year: 2000
Language: English
Pages: 852-856
Publication/Series: Nature Biotechnology
Volume: 18
Issue: 8
Document type: Journal article
Publisher: Nature Publishing Group

Abstract english

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.

Keywords

  • Antibody engineering
  • Diversity
  • In vitro evolution
  • Recombination

Other

Published
  • ISSN: 1087-0156
Carl Borrebaeck
E-mail: carl [dot] borrebaeck [at] immun [dot] lth [dot] se

Professor

Department of Immunotechnology

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