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

Research output: Contribution to journalArticle

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 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.

Details

Authors
Organisations
External organisations
  • BioInvent Therapeutic AB
  • Tohoku University
Research areas and keywords

Keywords

  • Antibody engineering, Diversity, In vitro evolution, Recombination
Original languageEnglish
Pages (from-to)852-856
Number of pages5
JournalNature Biotechnology
Volume18
Issue number8
Publication statusPublished - 2000
Publication categoryResearch
Peer-reviewedYes