A Solid-Phase Enzyme-Linked Immunospot (ELISPOT) Assay for Enumeration of Specific Antibody-Secreting Cells

J Immunol Methods. 1983 Dec 16;65(1-2):109-21. doi: 10.1016/0022-1759(83)90308-3.

Abstract

A solid-phase enzyme-linked immunosorbent assay (ELISPOT) is described for enumeration of cells secreting specific antibody. Spleen cells from immunized mice are incubated in antigen-coated polystyrene plates. After removal of the cells, bound antibodies are demonstrated by means of an immunoenzyme procedure in which enzyme-substrate reactions are performed in agarose. Dark-brown circular zones (spots), localized in areas of the dish where antibody production has occurred, are enumerated with the naked eye. Spectrophotometric estimation of enzyme-bound activity may be performed by substituting the gel for a liquid buffer, allowing accurate estimation of the total amount of secreted antibody. Versatile, sensitive and very easy to perform, this new assay provides a useful alternative to conventional plaque-forming cell assays.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity
  • Antibody-Producing Cells* / immunology
  • Antigens / immunology
  • Cell Count
  • Culture Media
  • Enzyme-Linked Immunosorbent Assay*
  • Female
  • Fetal Blood / immunology
  • Hemocyanins*
  • Hemolytic Plaque Technique
  • Immunoenzyme Techniques*
  • Kinetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Ovalbumin / immunology
  • Spleen / cytology
  • Spleen / immunology

Substances

  • Antigens
  • Culture Media
  • Ovalbumin
  • Hemocyanins
  • keyhole-limpet hemocyanin