Antibody engineering using phage display with a coiled-coil heterodimeric Fv antibody fragment.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 21552519.
- Also identified by DOI 10.1371/journal.pone.0019023 and PMC identifier 3084267.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
A Fab-like antibody binding unit, ccFv, in which a pair of heterodimeric coiled-coil domains was fused to V(H) and V(L) for Fv stabilization, was constructed for an anti-VEGF antibody. The anti-VEGF ccFv showed the same binding affinity as scFv but significantly improved stability and phage display level. Furthermore, phage display libraries in the ccFv format were constructed for humanization and affinity maturation of the anti-VEGF antibody. A panel of V(H) frameworks and V(H)-CDR3 variants, with a significant improvement in affinity and expressibility in both E. coli and yeast systems, was isolated from the ccFv phage libraries. These results demonstrate the potential application of the ccFv antibody format in antibody engineering.
Medical subject headings
- Immunoglobulin Fab Fragments
- Peptide Library
- Protein Engineering
- Protein Multimerization