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ELISpot and FluoroSpot: Antigen Specific Response Assays

ELISpot and FluoroSpot are highly sensitive functional assays that can be used throughout preclinical and clinical development, including phase 3 clinical trials
These assays can be extremely useful for measuring B cells, monocytes, dendritic cells, and antigen-specific T cells, which typically occur at low frequency in vivo. Both assays measure the cell-mediated immune response with accurate ex vivo frequency measurement of antigen-responsive cells down to the 1-in-100,000 cell range. Precision offers both ELISpot and FluoroSpot assays to support global preclinical and clinical projects, providing pre-validation and validation services using standard and custom assays.

Evaluating cell-mediated immune responses

We support global preclinical research with FluoroSpot and ELISpot testing—measuring cytokine-secreting cells at the single-cell level

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ELISpot assays

ELISpot assays provide robust measurement of cytokine-secreting cells at the single cell level. Like an ELISA, antibodies for the target cytokine are coated onto a 96-well plate. The cells being assessed are cultured in the same wells, stimulated to release cytokine, and then removed. Bound cytokines are visualized as with an ELISA, using chemiluminescence, with each spot corresponding to a single secreting cell.

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FluoroSpot assays

FluoroSpot assays are similar to ELISpot assays but rely on detection using fluorescently labeled detection antibodies.

Fluorescent detection enables simultaneous measurement of cells secreting different cytokines through multiplexing.

Example ELISpot assay panels

StimuliCell Type SimulatedDetailsStimulates Secretion of Human
CEF peptide pool CD8+ T cells Ag-specific stimulation by T cell epitopes derived from CMV, EBV, and influenza IFN-γ, IL-2, Granzyme B, Perforin
CEFT peptide pool CD4+ T cells Ag-specific stimulation by T cell epitopes derived from CMV, EBV, Tetanus Toxin and influenza IFN-γ, IL-2, IL-5, IL-13
Anti-CD3 Ab T cells Polyclonal T cell activation IFN-γ, 2, 4, 5, 10, 13, 17A
PHA T cells Polyclonal T cell activation IFN-γ, IL-2, 4, 5, 6, 10, 13, 17A, 21, GM-CSF, Granzyme B, Perforin
R848 + IL-2 B cells Polyclonal B cell activation IgG, IgA, IgM
Anti-CD40 + IL-4 B cells Polyclonal B cell activation IgE
LPS Monocytes/DC TLR4 agonist IL,1β, 6, 8, 10, 12, 23, GM-CSF, MIP1β, TNF-α
R848 Monocytes/DC TLR/8 agonist IFN-α, IL-1β, 6, 23, GM-CSF, TNF-α

Choosing the right immune monitoring technology

Understand use cases for cytokine profiling, cell-based immunophenotyping, gene expression, sequencing, circulating tumor markers, and immunohistochemistry and multiplex immunofluorescence.

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Guide to Immune Monitoring

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Guide to Immune Monitoring

Integrating biomarker and clinical trial data

To help researchers understand and interpret biomarker assay data in the context of other assays as well as the clinical trial data, Precision provides the option of managing and analyzing data using our proprietary QuartzBio Translational Informatics platform.

Learn more about the sample management, informatics, and computational capabilities of QuartzBio.

Global reach, multi-site support

Our FluoroSpot and ELISpot testing services support pre-clinical and clinical research, including multi-site studies, conducted anywhere in the world.

Working with Precision

Precision’s specialty lab scientists take a collaborative and consultative approach to projects and can provide recommendations on biomarker assay strategies and implementation. Services can be provided individually or as part of a comprehensive therapeutic development package including biomarker assays and clinical trials.

Additional lab services

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    Flow Cytometry

    Standard and spectral flow cytometry, on both research-grade and CLIA-validated instruments
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    Epiontis ID® Epigenetic Cell Profiling

    Precision’s proprietary immune monitoring technology which delivers robust, repeatable, and cost-effective immune cell phenotyping on any sample type
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    Cytokine Profiling & Proteomics

    Customized assays to detect cytokines or proteins of interest utilizing ELISA, Luminex, MSD, SIMOA, and Western Blotting

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