Human single-chain Fv (scFv) antibody specific to human IL-6 with the inhibitory activity on IL-6-signaling

Article type: Research Article

Authors: Gejima, R.^a | Tanaka, K.^a | Nakashima, T.^b | Hashiguchi, S.^a | Ito, Y.^a | Yoshizaki, K.^c | Sugimura, K.^{a; *}

Affiliations: [a] Department of Bioengineering, Faculty of Engineering, Kagoshima University, Kagoshima, Japan | [b] The Chemo-Sero-Therapeutic Research Institute, Kumamoto, Japan | [c] Department of Medical Science, Osaka University, Osaka, Japan

Correspondence: [*] Corresponding author: K. Sugimura, Professor, Dept. Bioengin', Facult. Engin', Kagoshima Univ., 1-21-40 Korimoto, Kagoshima 890-0065, Japan. Tel.: +81 99 285 8345; Fax: +81 99 2584706; E-mail: [email protected].

Abstract: 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 (Vγ-Vκ, Vγ-Vλ, Vμ-Vκ or μ-Vλ 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 (Vγ-Vλ), 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.

Keywords: ScFv, human antibody, IL-6, phage library, autoimmune diseases

DOI: 10.3233/HAB-2002-11402

Journal: Human Antibodies, vol. 11, no. 4, pp. 121-129, 2002