caa a22 4500 397556535 CHVBK 20180308164804.0 cr unu---uuuuu 161202e199609 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021462 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021462 Cloning and Sequencing of the VH and YK Genes of an Anti-CD3 Monoclonal Antibody, and Construction of a Mouse/Human Chimeric Antibody [Elektronische Daten] [Fumiko Arakawa, Masahide Kuroki, Motohisa Kuwahara, Tarumi Senba, Hiroaki Ozaki, Yuji Matsuoka, Yoshio Misumi, Hidetoshi Kanda, Takeshi Watanabe] Mouse monoclonal antibodies against CD3 on human T lymphocytes have been used for therapy in organ-transplant patients as a potent immunosuppressive agent or for treatment of cancer as a potent T cell activating agent. However, an inherent problem in their in vivo application is the human anti-mouse antibody response. In this study, we cloned and sequenced the variable region genes of the heavy and light chains (VH and VK) of a mouse anti-human CD3 monoclonal antibody (OKT3) using the reverse transcription-polymerase chain reaction method. Then, we constructed a mouse/human chimeric antibody, designated as Ch OKT3, by fusing the OKT3 VH and VK genes to the human heavy and light chain constant region genes (Cγ1 and CK) derived from a human plasma cell leukemia line (ARH77), respectively. The chimeric gene constructs were sequentially co-transfected into mouse non-Ig-producing hybridoma cells (Sp2/0) by electroporation. The Ch OKT3 antibody thus prepared bound to human peripheral blood mononuclear cells and competitively inhibited the binding of the parental MAb OKT3 to the blood mononuclear cells, indicating that this chimeric antibody seems to be suitable for in vivo therapeutic approaches. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence CD3 nationallicence chimeric antibody nationallicence monoclonal antibody nationallicence OKT3 nationallicence Arakawa Fumiko First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Kuroki Masahide First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Kuwahara Motohisa First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Senba Tarumi First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Ozaki Hiroaki Department of Ophthalmology, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Matsuoka Yuji First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Misumi Yoshio Second Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut Kanda Hidetoshi Immunological Research Laboratory, Research and Development Division, Eiken Chemical Co., Ltd. Ohtawara, Tochigi 324 aut Watanabe Takeshi Department of Molecular Immunology, Medical Institute of Bioregulation, Kyushu University Fukuoka 812 aut The Journal of Biochemistry Oxford University Press 120/3(1996-09), 657-662 0021-924X 120:3<657 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021462 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021462 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Arakawa Fumiko First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Kuroki Masahide First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Kuwahara Motohisa First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Senba Tarumi First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Ozaki Hiroaki Department of Ophthalmology, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Matsuoka Yuji First Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Misumi Yoshio Second Department of Biochemistry, School of Medicine, Fukuoka University 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-80 aut NATIONALLICENCE 700 1- Kanda Hidetoshi Immunological Research Laboratory, Research and Development Division, Eiken Chemical Co., Ltd. Ohtawara, Tochigi 324 aut NATIONALLICENCE 700 1- Watanabe Takeshi Department of Molecular Immunology, Medical Institute of Bioregulation, Kyushu University Fukuoka 812 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/3(1996-09), 657-662 0021-924X 120:3<657 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford