High sensitivity ELISpot/FluoroSpot assay for immune response evaluation

ELISpot is a reference technique for the detection and quantification of cellular immune responses at the single-cell level, enabling sensitive measurement of cytokine secretion from antigen-responsive immune cells. Its ability to detect rare but functionally active cells makes it highly relevant for immuno-oncology, vaccine development, and infectious disease research.

FluoroSpot extends these capabilities by allowing the simultaneous detection of multiple cytokines, providing deeper insights into immune cell polyfunctionality while reducing sample consumption.

This application note presents key workflow considerations, assay optimisation strategies, qualification data, and comparative performance assessment of ELISpot and FluoroSpot assays for immunogenicity testing and immune monitoring.

Supporting your immunogenicity assessment strategy

Optimised ELISpot and FluoroSpot workflows provide a robust framework for evaluating antigen-specific cellular immune responses across the drug development continuum. It:

• Supports sensitive detection of antigen-specific T-cell responses at the single-cell level
• Provides reproducible measurements through qualified assay conditions and demonstrated precision
• Facilitates comprehensive immune profiling through multiplex cytokine detection using FluoroSpot
• Improves confidence in the interpretation of low-frequency immune responses by reducing background signal
• Enables evaluation of both direct antigen stimulation and expanded memory responses following in vitro stimulation
• Supports immune monitoring activities in regulated environments using workflows compatible with fresh and cryopreserved PBMC samples

Together, these approaches provide a flexible platform for functional characterisation of cellular immune responses

Typical applications

• Immunogenicity assessment
• Vaccine development and vaccine immune monitoring studies
• Immuno-oncology research and immune response evaluation
• Infectious disease research and assessment of antigen-specific T-cell responses
• Characterisation of mechanisms of action and comparison of antigen formulations
• Monitoring durable memory T-cell responses during preclinical and clinical studies