title: Antibody engineering using phage display with a coiled-coil heterodimeric Fv antibody fragment.
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.
authors.affiliation: Biologics Discovery, Merck Research Laboratories, Merck & Co., Inc., Palo Alto, California, United States of America.
authors.forename: Xinwei
authors.initials: X
authors.lastname: Wang
authors.forename: Pinyu
authors.initials: P
authors.lastname: Zhong
authors.forename: Peter P
authors.initials: PP
authors.lastname: Luo
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D005240:Feasibility Studies; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics; D007167:Immunotherapy; D008969:Molecular Sequence Data; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods*; D055503:Protein Multimerization; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology
chemical_list: D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D042461:Vascular Endothelial Growth Factor A