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

Methods Mol Biol. 2019:1904:147-162. doi: 10.1007/978-1-4939-8958-4_6.

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.

Keywords: Antibody-secreting cell; Antigen-specific antibody; Immunospot array assay on a chip; Microwell-array chip.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Formation / genetics
  • Antibody Formation / immunology*
  • Antibody-Producing Cells / immunology*
  • Antibody-Producing Cells / metabolism
  • Antigens / immunology*
  • Biomarkers
  • Epitopes / immunology*
  • Gene Expression
  • Genetic Vectors / genetics
  • Hybridomas / immunology
  • Immunoassay* / methods
  • Immunophenotyping
  • Leukocytes, Mononuclear / immunology
  • Leukocytes, Mononuclear / metabolism
  • Microarray Analysis* / methods

Substances

  • Antigens
  • Biomarkers
  • Epitopes