title: Optimization of anti-tumor necrosis factor-alpha single chain Fv displayed on phages for creation of functional antibodies.
abstract: In this study, we converted the immunoglobulin-type anti-human tumor necrosis factor-alpha (TNF-alpha) monoclonal antibody (Mab) to a scFv-type antibody in order to assess its basic properties. The immunoglobulin VH and VL genes were isolated from the hybridoma that produced an anti-TNF-alpha neutralizing Mab, and they were then linked together to create scFvs of the VL-VH or VH-VL-form. The binding affinity to TNF-a was retained in both scFvs. Interestingly, the VL-VH-type scFv effectively inhibited the TNF-alpha-mediated cytotoxicity, while this neutralization activity was dramatically decreased in the VH-VL-type scFv. These results suggest that the VL-VH-type scFv is a suitable template to create improved versions of the anti-TNF-alpha antibody using a phage display system, and they also show that the structural format must be taken into account in manufacturing scFvs.
authors.affiliation: Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation, Ibaraki, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Mukai
authors.forename: T
authors.initials: T
authors.lastname: Okamoto
authors.forename: M
authors.initials: M
authors.lastname: Kawamura
authors.forename: H
authors.initials: H
authors.lastname: Shibata
authors.forename: T
authors.initials: T
authors.lastname: Sugita
authors.forename: S
authors.initials: S
authors.lastname: Imai
authors.forename: Y
authors.initials: Y
authors.lastname: Abe
authors.forename: K
authors.initials: K
authors.lastname: Nagano
authors.forename: T
authors.initials: T
authors.lastname: Nomura
authors.forename: H
authors.initials: H
authors.lastname: Kamada
authors.forename: Y
authors.initials: Y
authors.lastname: Tsutsumi
authors.forename: T
authors.initials: T
authors.lastname: Mayumi
authors.forename: S
authors.initials: S
authors.lastname: Nakagawa
authors.forename: S
authors.initials: S
authors.lastname: Tsunoda
mesh_terms: D019138:Antibodies, Blocking / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D019151:Peptide Library; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D014409:Tumor Necrosis Factor-alpha