Determination of the antigen/epitope that is recognized by human monoclonal antibody CLN-IgG.
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.
Hagiwara
H
H
Hagiwara Institute of Health, Kasai-City, Hyogo, Japan. info@hih.org
Aotsuka
Y
Y
Yamamoto
Y
Y
Miyahara
J
J
Mitoh
Y
Y
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D001483:Base Sequence; D001666:Binding Sites, Antibody; D001932:Brain Neoplasms / Q000276:immunology / Q000473:pathology; D002846:Chromatography, Affinity; D004247:DNA; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D005909:Glioblastoma / Q000276:immunology / Q000473:pathology; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes; D014746:Vimentin; D004247:DNA