Analysis of cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulin(METHODS)
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概要
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Although various in vitro immunization methods to generate antigen-specific antibodies have been described, a highly effective method that can generate high-affinity immunoglobulins has not yet been reported. Herein, we analyzed a cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulins. We identified a combination of T cell-dependent stimuli (IL-4, IL-5, anti-CD38 and anti-CD4O antibodies) plus lipopolysaccharides (LPS) that stimulates antigen-exposed splenocytes in vitro, followed by induction of the cells phenotypically equivalent to germinal center B cells. We also observed that LPS induced high expression levels of mRNA for activation-induced cytidine deaminase. We stimulated antigen-exposed splenocytes, followed by the accumulation of mutations in immunoglobulin genes. From the immunized splenocytes, hybridoma clones secreting antigen-specific immunoglobulins were obtained.
- 公益社団法人日本生物工学会の論文
- 2013-03-00
著者
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Nagamune Teruyuki
Department Of Bioengineering Center For Nanobio Integration The University Of Tokyo:department Of Ch
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Kawahara Masahiro
Department Of Analytical Chemistry Faculty Of Pharmaceutical Sciences The University Of Tokyo
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Kawahara Masahiro
Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo
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Yoshimi Tatsunari
Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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Inagaki Takashi
Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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Kobayashi Satoshi
Innovative Antibody Engineering Laboratory, Advance Co. Ltd.
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Nagamune Teruyuki
Department of Bioengineering, Graduate School of Engineering, University of Tokyo:Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo
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