Highly Sensitive Assays on ELISpot and FluoroSpot to Measure Cytokine Secreting Cells

Detecting cytokines at the single-cell level

Enzyme-Linked Immunosorbent Spot (ELISpot) and FluoroSpot build on familiar immunoassay principles to assess the frequency of antigen-specific cytokine-secreting cells at a single cell level.

With its high sensitivity, it can detect low antigen-specific responses and be used to complement information from other functional assays, including intracellular cytokine staining (ICS) by flow cytometry and ELISA.

ELISpot and FluoroSpot assays are used to characterize pre-existing immune responses, monitor the immunogenicity of vaccines, and assess the effects of immune-modulating therapies in fields such as cancer, infectious diseases, autoimmune and rare/orphan diseases.

Our scientists have extensive experience in customizing and validating ELISpot assays to specific antigens for exploratory, secondary, or primary endpoint use in clinical trials.

What is ELISpot - IFNγ

ELISpot is an efficient assay that requires the use of a small number of cells, allowing for multiple stimulations, and necessitates minimal cell manipulations (no fixation/permeabilization).

It is also a highly sensitive method, usually showing sensitivity higher than an intra-cellular cytokine staining.

CellCarta also has expertise with the cultured ELISpot assay which allows the monitoring of responses by memory T cells.

Fit-for-purpose ELISpot assay for the detection of IFNγ

  • Readily available for exploratory endpoints and can be validated for secondary and primary endpoints
  • Validation performed on cryopreserved peripheral blood mononuclear cells (PBMC)
  • Standardization of the SFU count using ImmunoSpot® S6 Universal analyzer
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What is FluoroSpot

With FluoroSpot assays, analyte-secreting cells are detected using antibodies labeled with fluorophores instead of enzymes, resulting in a one-step direct detection as compared to the two-step detection system of ELISpot.

FluoroSpot also has the advantage of enabling the analysis of multiple analytes in one well.

The technology allows for the identification of cells producing each analyte as well as the ones producing both of these analytes.

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Customized Assays from CellCarta

Our experienced team can customize both ELISpot and FluoroSpot assays according to the antigen stimulation selected and the cell type investigated.

The well-established validation process we follow leads to optimized assays and guarantees confidence in your results.

Key Applications of ELISpot and FluoroSpot

  • Monitoring immunogenicity of vaccines, cell therapies, and immune-modulating therapies (CAR-T, checkpoint inhibitors, cytokines, agonists, and adjuvants)
  • Large-scale epitope mapping
  • Detection of HLA-restricted responses
  • Measuring the immune responses of memory B cells and T cells from PBMC
  • Assessing the immune response of T cells co-cultured with dendritic cells
  • Providing readouts on selected populations isolated with cell separation techniques such as magnetic separation columns