title: Generation of human monoclonal antibodies recognising membranous antigens of the lung adenocarcinoma cell line A549 using an AMeX immunohistostaining method.
abstract: Four monoclonal antibodies (MAbs) from hybridoma obtained by in vitro stimulation of regional lymph node lymphocytes from lung cancer patients and electrofusion of the stimulated cells with murine or human-mouse myeloma cells were reactive to lung cancer cells in enzyme-linked immunoabsorbent assay, and to lung cancer tissue in immunohistochemical analysis using acetone-methyl benzoate-xylene (AMeX) fixed tissue and in immunofluorescence analysis. Three of the MAbs (designated ZLG40, 27D57 and 28K29) recognised cell-surface antigens of the lung adenocarcinoma cell line A549 and the remaining one (designated 29D38) recognised nuclear membrane antigens of the same cell line. The three surface-binding MAbs showed a significant complement-dependent cytotoxicity (CDC) to the A549 cells, but the membrane-binding 29D38 showed no CDC to the A549 cells. Western blotting of the extracts of the A549 or PC6 (small-cell lung cancer) cell lines by the four MAbs showed a 28K29 antigen band at M(r) of approximately 600,000 (+/- 2-ME), a ZLG40 antigen band at M(r) 50,000 (+/- 2-ME), and one 29D38 antigen band at M(r) of more than 1,000,000 (-2-ME) and M(r) between 20,000 and 80,000 (+2-ME), but no detectable band for 27D57 antigen.
authors.affiliation: Diagnostics Research and Development Department, Asahi Chemical Industry Co. Ltd., Shizuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yoshinari
authors.forename: H
authors.initials: H
authors.lastname: Kimura
authors.forename: K
authors.initials: K
authors.lastname: Arai
authors.forename: I
authors.initials: I
authors.lastname: Sugawara
authors.forename: K
authors.initials: K
authors.lastname: Noda
authors.forename: M
authors.initials: M
authors.lastname: Kihara
authors.forename: H
authors.initials: H
authors.lastname: Misaki
authors.forename: Y
authors.initials: Y
authors.lastname: Yamaguchi
mesh_terms: D000230:Adenocarcinoma / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis; D003602:Cytotoxicity, Immunologic; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000276:immunology*; D051379:Mice; D008970:Molecular Weight; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface