Human single-chain Fv (scFv) antibody specific to human IL-6 with the inhibitory activity on IL-6-signaling.
Human anti-IL-6 antibody may be useful for the immunotherapy of various inflammatory diseases, such as rheumatoid arthritis. Since IL-6 is a growth factor for B cell hybridoma, it is not easy to isolate murine B cell hybridomas producing the anti-IL-6 antibody with the IL-6-signaling inhibitory activity. In this study, the antibody library (Vgamma-Vkappa, Vgamma-Vlambda, Vmu-Vkappa or micro-Vlambda ligated into the pCANTAB 5E phagemid vector) was prepared from peripheral blood mononuclear cells of 20 healthy subjects. The phage display library was panned with an IL-6-coated plastic plate, and the binding specificity was confirmed by ELISA and BIAcore. From the antibody library (Vgamma-Vlambda), five IL-6-specific phage clones were isolated. The effects of the soluble scFvs purified from these phage clones were tested on the growth of the IL-6-dependent human cell line, KT-3. Two of these clones significantly inhibited the growth of KT-3, and three showed no inhibition.
Gejima
R
R
Department of Bioengineering, Faculty of Engineering, Kagoshima University, Kagoshima, Japan.
Tanaka
K
K
Nakashima
T
T
Hashiguchi
S
S
Ito
Y
Y
Yoshizaki
K
K
Sugimura
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001327:Autoimmune Diseases / Q000628:therapy; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000097:blood / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region; D015850:Interleukin-6 / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008969:Molecular Sequence Data; D019151:Peptide Library; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000276:immunology*; D020349:Surface Plasmon Resonance
D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D015850:Interleukin-6; D019151:Peptide Library; C055966:immunoglobulin Fv