(Enzyme-Linked ImmunoSpot)

For Qualitative Analysis of Cellular Immune Responses

BioAgilytix offers Enzyme-Linked ImmunoSpot (ELISpot) assays at our European laboratory in Hamburg, Germany. Qualified and validated for regulated work, BioAgilytix’s ELISpot platform provides robust data on cell-mediated immunity for regulated immune monitoring studies for biologics, Cell and Gene Clinical Development Support.

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How the ELISpot Platform Works

ELISpot assays employ the sandwich enzyme-linked immunosorbent assay (ELISA) technique, where either a monoclonal or polyclonal antibody specific for the chosen analyte is pre-coated onto a PVDF (polyvinylidene difluoride)-backed microplate. Freshly isolated, thawed, or cultured cells are plated together with the antigens of interest and incubated to allow for the activation of the antigen-specific T cells and the induction of their cytokine secretion.
As the antigen-specific cells are activated, they release the cytokine, which is captured directly on the membrane surface by the immobilized antibody. The visualization of the cytokines secreted individual T- or B-clones is achieved by enzyme-conjugated or fluorescence-based substrates. The fluorescence-based method allows a multiplex analysis of antigen-specific clones. The spots are then counted with the automated ELISpot reader system.

Why BioAgilytix for Small and Large Molecule Bioanalysis?

Our passion for the science of small and large molecule bioanalysis drives our ability to deliver premium, tailored services for biomarker, immunogenicity, pharmacokinetics (PK), and cell-based assays.


The ELISpot platform is considered among the most useful for determining cell-mediated immunity because of its sensitivity and specificity in detecting rare antigen-specific T cells (or B cells). ELISpot assays are able to provide qualitative information regarding the specific cytokine or other secreted immune molecule.

We use ELISpot to visualize the secretory product(s) of individual activated or responding cells to analyze the frequencies of rare antigen-specific cells within a test population, such as single positive cells within a population of peripheral blood mononuclear cells (PBMCs).

At BioAgilytix, we most frequently utilize ELISpot in our work with Cytotoxic T Cell Response, Helper T Cell Response, B Cell Antibody response, Epitope Discovery, immune response monitoring, epitope discovery, and helper T cell response evaluation. Specific to bioanalytical assessment of cell and gene therapies, ELISpot can be used to assess cellular immune responses against the vector. We can also use fluorescently labeled antibodies, even in parallel via multiplexing.


ELISpot offers exceptional sensitivity for the detection and enumeration of rare antigen-specific cells because of its method of capturing and secreting cytokine, which overcomes some of the limitations of the traditional ELISA method. ELISpot allows for detection of a single cell that secrets a protein of interest (cytokine, effector protein, receptor, surface marker, antibody) – and since the cytokine released in response to antigen can be mapped to a single cell, T cell responder frequencies can be calculated.

It also provides an indication of the type of cytokine response that has been elicited and hence the type of immune response. As such, ELISpot assays are able to provide high-quality qualitative information regarding the specific cytokine or other secreted immune molecule.

Top Benefits of ELISpot Assays:

  • High Sensitivity
  • Single Cell Detection and Enumeration
  • Scalability for Higher Throughput
  • Multiplexing of Analytes

Platform In Action:

Managing unwanted immunogenicity of therapeutic proteins

Learn why the European Immunogenicity Platform (EIP) believes that a multidisciplinary approach is warranted to better understand and minimize drug immunogenicity.


With ELISpot, our scientific team is able to achieve the degree of sensitivity needed to accurately detect and enumerate rare antigen-specific cells and evaluate helper T cell response.

In fact, BioAgilytix’s ELISpot platform is qualified and validated for regulated studies involving biologics and gene therapies, and our team has a proven track record of leveraging this platform to deliver high-quality qualitative results.

Speak with one of our ELISpot experts about your scientific requirements, and we will discuss how the features of this platform can be optimally applied your assay requirements. ELISpot experts available in both our US and European labs.

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