The enzyme-linked immunospot (ELISPOT) assay is a powerful tool for detecting
and enumerating individual cells that secrete a particular protein in vitro.1 Based
on the sandwich enzyme-linked immunosorbent assay (ELISA), the ELISPOT assay
derives its specificity and sensitivity by employing high affinity capture and detection
antibodies and enzyme-amplification. Although originally developed for analyzing
specific antibody-secreting cells,2,3 the assay has been adapted for measuring the
frequencies of cells that produce and secrete other effector molecules, such as
cytokines.4,5,6 The sensitivity of the assay lends itself to measurement of even very
low frequencies of analyte-producing cells (eg, 1/300,000).1 Recent developments
in assay plate design and in ELISPOT plate-reader instrumentation have significantly
improved the utility of the ELISPOT method for objective and rapid analysis of
analyte-producing cells.
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