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

Summary, in 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.

• Författare: 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
• Avdelning/ar: Institutionen för immunteknologi
• Publiceringsår: 2000
• Språk: Engelska
• Sidor: 852-856
• Publikation/Tidskrift/Serie: Nature Biotechnology
• Volym: 18
• Issue: 8
• Länkar: http://dx.doi.org/10.1038/78458
• Dokumenttyp: Artikel i tidskrift
• Förlag: Nature Publishing Group
• Nyckelord: Antibody engineering; Diversity; In vitro evolution; Recombination
• Status: Published
• ISBN/ISSN/Övrigt: ISSN: 1087-0156