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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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Enzyme-Linked ImmunoSpot (ELISpot) for Single-Cell Analysis.

Sylvia Janetzki1, Rachel Rabin2

  • 1ZellNet Consulting, Inc., 555 North Avenue, Suite 25-S, Fort Lee, NJ, 07024, USA. sylvia@zellnet.com.

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|November 7, 2015
PubMed
Summary

The Enzyme-Linked ImmunoSpot (ELISpot) assay detects low-abundance analytes by capturing secreted molecules from single immune cells. This robust technique quantifies cellular responses and is expanding to analyze polyfunctional cells.

Keywords:
B cell assaysELISpotFluoroSpotImmune monitoringT cell assays

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Area of Science:

  • Immunology
  • Biotechnology
  • Cellular Biology

Background:

  • The Enzyme-Linked ImmunoSpot (ELISpot) is a heterogeneous immunoassay crucial for detecting low-abundance analytes.
  • It enables monitoring of immune system responses at the single-cell level by capturing secreted molecules.

Purpose of the Study:

  • To describe optimized protocol steps for the ELISpot technique.
  • To highlight improvements and tools for the scientific community.
  • To discuss the expansion of ELISpot into polyfunctional cell analysis.

Main Methods:

  • Utilizes membrane-bound antibodies to capture secreted molecules from individual cells.
  • Employs ELISA-related development steps to visualize captured molecules.
  • Generates distinct spots representing cytokine-secreting cells for quantification.

Main Results:

  • Provides quantification of cells secreting specific molecules.
  • Offers insights into the kinetics and strength of cellular secretion.
  • Demonstrates the adaptability of ELISpot for advanced cellular analysis.

Conclusions:

  • The ELISpot assay is a reliable and robust method for single-cell immune monitoring.
  • Optimized protocols and available tools enhance its utility.
  • The technique's expansion into polyfunctional analysis broadens its applications in immunology research.