Lung cancer-reacting human recombinant antibody AE6F4: Potential usefulness in the sputum cytodiagnosis

Article type: Research Article

Authors: Shoji, Masahiro^{a; *} | Kawamoto, Seiji^b | Seki, Kiyohiko^c | Teruya, Kiichiro^b | Setoguchi, Yuko^a | Mochizuki, Katsumi^a | Kato, Masatoshi^a | Hashizume, Shuichi^a | Hanagiri, Takeshi^d | Yoshimatsu, Takashi^d | Nakanishi, Kozo^d | Yasumoto, Kosei^d | Nagashima, Akira^e | Nakahashi, Hisashi^f | Suzuki, Tatsuo^g | Imai, Tadaaki^h | Shirahata, Sanetaka^b | Nomoto, Kikuoⁱ | Murakami, Hiroki^b

Affiliations: [a] Morinaga Institute of Biological Science, 2-1-1 Shimosueyoshi, Tsurumi-ku, Yokohama 230, Japan | [b] Graduate School of Genetic Resources Technology, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812, Japan | [c] Department of Applied Biological Sciences, Faculty of Agriculture, Saga University, 1 Honjho-cho, Saga 840, Japan | [d] Second Department of Surgery, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807, Japan | [e] Department of Chest Surgery, Kitakyushu Municipal Medical Center, 2-1-1 Bashaku, Kokurakita-ku, Kitakyushu 802, Japan | [f] Matsuyama Red Cross Hospital, 1 Bunkyo-cho, Matsuyama 790, Japan | [g] Department of Research Planning, Kitasato Institute Hospital, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan | [h] Section of Pathology, The Kitasato Institute Medical Center Hospital, 121-1 Arai, Kitamoto, Saitama 364, Japan | [i] Department of Immunology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku Fukuoka 812, Japan

Correspondence: [*] Correspondence and reprint requests to: Masahiro Shoji, Morinaga Institute of Biological Science, 2-1-1 Shimosueyoshi, Tsurumi-ku, Yokohama 230, Japan.

Abstract: Human monoclonal antibody (hMAb) AE6F4 has been shown to be potentially useful for immunocytological detection of lung cancer cells in sputum. By recombinant DNA technology, IgM type hMAb AE6F4 was switched to IgG. The IgG mimic recombinant AE6F4 antibody expression plasmid was assembled using the antibody heavy chain gene, which ligated the gene encoding VH and CH1 (μ) domains of hMAb AE6F4 heavy chain to the gene encoding CH2(γ 1) and CH3(γ1) domains of human IgG heavy chain, and the antibody light chain gene of hMAb AE6F4. The recombinant antibody expressed by baby hamster kidney (BHK) – 21 cells showed molecular size equivalence to IgG, and consisted of human μ–γ hybrid heavy and κ light chains. The immunological specificity of the recombinant antibody was the same as that of hMAb AE6F4 by immunoblotting analysis to the 14–3–3 protein, the putative antigen of hMAb AE6F4, and by immunohistochemical and immunocytological analyses using tissue sections and sputa of lung cancer patients. The transfected BHK-21 cells produced the recombinant antibody persistently and the productivity was greater than 20 times that by human-human hybridoma producing hMAb AE6F4.

Keywords: Human monoclonal antibody, recombinant antibody, lung cancer, baby hamster kidney cells, diagnostics

DOI: 10.3233/HAB-1996-7104

Journal: Human Antibodies, vol. 7, no. 1, pp. 27-36, 1996