A rapid and efficient strategy to generate antigen-specific human monoclonal antibody by in vitro immunization and the phage display method

Shin ei Matsumoto, Makiko Yamashita, Yoshinori Katakura, Yoshihiro Aiba, Kosuke Tomimatsu, Shigeru Kabayama, Kiichiro Teruya, Sanetaka Shirahata

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

An in vitro immunization (IVI) protocol was developed for inducing antigen-specific immune responses in human peripheral blood mononuclear cells (PBMCs). After antigen sensitization of PBMCs by IVI, B cells producing antigen-specific antibody can be propagated within a week. Here, we attempted to establish a rapid, efficient strategy to obtain antigen-specific antibody by the phage display method using in vitro immunized PBMCs. Heavy and light chain variable region genes were easily amplified from these PBMCs immunized with mite extract (ME). After generating a combinatorial phage library (1.6 × 105 members), 4 antigen-specific clones were selected by 5 panning rounds using biotinylated antigen and streptavidin magnetic beads. Next, we combined variable region genes of these selected clones with human IgG constant region genes and produced human IgG-type antibody. Direct and competitive enzyme-linked immunosorbent assays demonstrated that the mAb 1C11 clone bound specifically to ME. We thus established a rapid, efficient method to obtain antigen-specific human antibody genes and produce human monoclonal IgG antibody using the phage antibody library generated from in vitro immunized PBMCs.

Original languageEnglish
Pages (from-to)2-9
Number of pages8
JournalJournal of Immunological Methods
Volume332
Issue number1-2
DOIs
Publication statusPublished - Mar 20 2008

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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