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Isolation of Antigen-Specific, Antibody-Secreting Cells Using a Chip-Based Immunospot Array

  • Hiroyuki KishiEmail author
  • Tatsuhiko Ozawa
  • Hiroshi Hamana
  • Eiji Kobayashi
  • Atsushi Muraguchi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1904)

Abstract

Antigen-specific monoclonal antibodies are useful tools to detect very small amounts of antigenic materials and are applicable for antibody therapeutics. To produce mouse monoclonal antibodies, a hybridoma between B lymphocytes and myeloma cells is used to produce antigen-specific monoclonal antibodies. However, a good hybridoma system is not available to obtain human monoclonal antibodies. To produce antigen-specific human monoclonal antibodies, transformation of B lymphocytes with Epstein-Barr viruses or a phage-display system is used. Here, we describe the screening of antigen-specific, antibody-secreting cells using microwell array chips to obtain antigen-specific human monoclonal antibodies. The system can be applied to screen antigen-specific, antibody-secreting cells from any animal species.

Key words

Antigen-specific antibody Antibody-secreting cell Microwell-array chip Immunospot array assay on a chip 

Notes

Acknowledgment

This work was supported by grants from the Toyama Medical Bio-Cluster Project and Hokuriku Innovation Cluster for Health Science of the Ministry of Education, Culture, Sports, Science and Technology, Japan. This work was also supported by JSPS KAKENHI Grant Number JP16H06499 (H.K.), the Platform Project for Supporting in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from the Ministry of Education, Culture, Sports, Science (MEXT) and the Japan Agency for Medical Research and Development (AMED) (A.M.), and the Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from AMED under Grant Number JP18am0101077 (T.O.).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Hiroyuki Kishi
    • 1
    Email author
  • Tatsuhiko Ozawa
    • 1
  • Hiroshi Hamana
    • 1
  • Eiji Kobayashi
    • 1
  • Atsushi Muraguchi
    • 1
  1. 1.Department of Immunology, Graduate School of Medicine and Pharmaceutical SciencesUniversity of ToyamaToyamaJapan

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