Optimization of anti-tumor necrosis factor-alpha single chain Fv displayed on phages for creation of functional antibodies.
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.
Mukai
Y
Y
Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation, Ibaraki, Japan.
Okamoto
T
T
Kawamura
M
M
Shibata
H
H
Sugita
T
T
Imai
S
S
Abe
Y
Y
Nagano
K
K
Nomura
T
T
Kamada
H
H
Tsutsumi
Y
Y
Mayumi
T
T
Nakagawa
S
S
Tsunoda
S
S
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
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D014409:Tumor Necrosis Factor-alpha