Production of Human Monoclonal Antibodies against FcεRIα by a Method Combining in Vitro Immunization with Phage Display
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概要
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An in vitro immunization protocol using human peripheral blood mononuclear cells (PBMC) was developed to generate human antigen-specific antibodies. Monoclonal antibodies have great potential, and in particular, efficient acquirement of monoclonal antibodies against membrane proteins provides advantages. In this study, we tried to generate a human monoclonal antibody against the high affinity IgE receptor, FcεRIα, using a method combining in vitro immunization and phage display. Heavy and light chain variable region genes were obtained from PBMC immunized in vitro with FcεRIα-expressed KU812F cells. Subsequently a combined phage antibody library 6×103 in the size was generated. Antigen-specific phage antibody clones were selected by panning with recombinant FcεRIα and recombined to produce human IgG format antibodies using CHO cells. The antibodies exhibited specific binding against FcεRIα. These results suggest that one can obtain membrane protein-specific human monoclonal antibodies from a relatively small phage antibody library using in vitro immunized PBMCs.
- 社団法人 日本農芸化学会の論文
著者
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SHIRAHATA Sanetaka
Graduate School of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
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TERUYA Kiichiro
Graduate School of Systems Life Sciences, Kyushu University
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KABAYAMA Shigeru
Nihon Trim Co., Ltd.
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KATAKURA Yoshinori
Graduate School of Systems Life Sciences, Kyushu University
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Tomimatsu Kosuke
Graduate School of Systems Life Sciences, Faculty of Agriculture, Kyushu University
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YAMASHITA Makiko
Graduate School of Systems Life Sciences, Faculty of Agriculture, Kyushu University
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MATSUMOTO Shin-ei
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University
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Tomimatsu Kosuke
Graduate School Of Systems Life Sciences Kyushu Univ.
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