Determination of the antigen/epitope that is recognized by human monoclonal antibody CLN-IgG

Article type: Research Article

Authors: Hagiwara, Hideaki^* | Aotsuka, Yasuyuki | Yamamoto, Yasushi | Miyahara, Junichi | Mitoh, Yuko

Correspondence: [*] Corresponding author: Hideaki Hagiwara, Hagiwara Institute of Health, 1173 Asazuma-cho, Kasai-City, Hyogo, 679-0105 Japan. Tel.: +81 790 47 1581; Fax: +81 790 47 1585; E-mail: info@hih.org.

Abstract: Ever since the development of human monoclonal antibody CLN-IgG in 1982, we anticipated the identification of the antigen that is recognized by this antibody. Despite its scarce expression on the cell surface, susceptibility to proteolytic enzymes and adherence to experimental equipment, we finally succeeded in determining the antigen moiety that is recognized by this antibody by means of CLN-IgG conjugated column affinity chromatography, two-dimensional electrophoresis, MALDI-TOF/MS and use of glioblastoma cell line U-251MG. The antigen was found to be vimentin, a cytoskeletal protein, and we succeeded in determining a 79 amino acids sequence of the epitope which turned out to comprise a part of the c2 (coil 2 of the central rod) domain of vimentin.

Keywords: human monoclonal antibody, vimentin, tumor antigen

DOI: 10.3233/HAB-2001-10204

Journal: Human Antibodies, vol. 10, no. 2, pp. 77-82, 2001