In the Zone: ELISpot assays for evaluating safety
A critical aspect of testing therapeutic drugs that treat human disease is evaluating their safety post-administration. Traditionally, this has been achieved by monitoring key biomarkers, which are biological indicators of disease or patient condition, and through bioanalytical testing of patient samples for an anti-drug immunologic response or immunogenicity. Immunogenicity is measured using assays that detect the humoral immune response, i.e. antibodies that bind to the drug. However, a complete immune response consists of not only the antibodies produced by B cells but also the activation of T cells, which is referred to as cellular immunogenicity. Monitoring cellular immunogenicity is a recent requirement for therapeutics such as chimeric antigen receptors and viral vector-based therapies. Enzyme-linked immunosorbent spot assays, or ELISpot, is one technology being applied to such testing strategies. For this method, the drug’s amino acid sequence is used to design overlapping short peptides, which are added to a sample of a patient’s immune cells or peripheral blood mononuclear cells in vitro. Activation of T cells in response to the peptides is then detected through quantitating the number of cells that produce inflammatory cytokines after stimulation. ELISpot assays have become a common aspect of safety testing for cell and gene therapies in both preclinical and clinical settings and is a highly sensitive tool for measuring immune responses.
INTERVIEW
An overview of the challenges and implementation of ELISpots: an interview with Michelle Miller
In this interview, Michelle Miller (BioAgilytix; NC, USA) provides a basic overview of an ELISpot assay and how the method is used for the safety testing of therapeutics. She discusses some of the technical challenges with using ELISpots in regulated bioanalysis including the appropriate handling of clinical samples, quick isolation of peripheral blood mononuclear cells after sample collection and sample logistics including proper control of temperature. She also reveals her hope for more specific guidance on the use of ELISpot assays for regulated bioanalysis.
INFOGRAPHIC
Infographic: the use of ELISpot assays for evaluating the safety of therapeutic drugs
After dosing with a drug, patient immune systems can mount anti-drug responses, which include humoral and cellular immunogenicity. Explore this ELISpot assay workflow to discover more about current guidelines and some common areas of concern.
RESOURCES
Recommendations on ELISpot assay validation by the GCC
Bioanalytical challenges for cellular immunogenicity testing using validated ELISpot assays
ELISpot method development and validation for gene therapies
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