Production of human monoclonal antibodies against FcεRIα by a method combining in vitro immunization with phage display

Kosuke Tomimatsu, Shin Ei Matsumoto, Makiko Yamashita, Kiichiro Teruya, Yoshinori Katakura, Shigeru Kabayama, Sanetaka Shirahata

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1465-1469
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number7
DOIs
Publication statusPublished - Aug 19 2009

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Immunization
Bacteriophages
Display devices
Monoclonal Antibodies
Antibodies
Blood Cells
Membrane Proteins
Blood
IgE Receptors
Antigens
CHO Cells
In Vitro Techniques
Clone Cells
Immunoglobulin G
Genes
Light

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Production of human monoclonal antibodies against FcεRIα by a method combining in vitro immunization with phage display. / Tomimatsu, Kosuke; Matsumoto, Shin Ei; Yamashita, Makiko; Teruya, Kiichiro; Katakura, Yoshinori; Kabayama, Shigeru; Shirahata, Sanetaka.

In: Bioscience, Biotechnology and Biochemistry, Vol. 73, No. 7, 19.08.2009, p. 1465-1469.

Research output: Contribution to journalArticle

Tomimatsu, Kosuke ; Matsumoto, Shin Ei ; Yamashita, Makiko ; Teruya, Kiichiro ; Katakura, Yoshinori ; Kabayama, Shigeru ; Shirahata, Sanetaka. / Production of human monoclonal antibodies against FcεRIα by a method combining in vitro immunization with phage display. In: Bioscience, Biotechnology and Biochemistry. 2009 ; Vol. 73, No. 7. pp. 1465-1469.
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