EpiScreen Time Course T Cell Assay | Abzena

EpiScreen™ Time Course T Cell Assay

EpiScreen™ is an accurate and sensitive way to assess the potential immunogenicity of proteins and antibodies ex vivo by measuring CD4+ T cell responses, the primary drivers of memory-based immunogenicity. The EpiScreen™ time course T cell assay is used extensively across the industry as a standard for preclinical immunogenicity testing.

The assay provides an accurate and sensitive way to assess the overall immunogenicity of protein therapeutics during early lead selection or during manufacturing. For each study a donor cohort is selected to best represent the distribution of HLA-allotypes in the human population.

By measuring CD4+ T cell responses ex vivo, the assay assesses the strength and kinetics of an individual’s T cell response to a protein therapeutic, thereby giving an indication of an immunogenic response against the protein.

The assay can facilitate lead selection based on the immunogenicity of whole proteins including mAbs and other human and non-human protein therapeutics. It can also assess the immunogenicity following antibody humanisation and protein deimmunisation.

About the assay

Assays are performed using bulk cultures of peripheral blood mononuclear cells (PBMCs), typically from 50 individual donors, with a distribution of HLA-DR allotypes (coverage and frequency) representing the human population of interest.
T cell proliferation and IL-2 secretion, both markers of T cell activation, are measured using 3H-thymidine incorporation and ELISpot, respectively. Strong correlation between these markers enables the detection of CD4+ effector T cell activation and discriminates between T cell effector and regulatory responses.

Right:  Healthy donor T cell proliferation responses to both A33 and Trastuzumab.  PBMC from bulk cultures were sampled and assessed for proliferation after incubation with Trastuzumab and A33.  Right top: Maxmimal proliferation responses with an stimulation Index ≥ 1.90 (indicated by the dotted line) that were significant (p < 0.05) using an unpaired, two sample student’s t test were considered positive.

Right bottom: The percentage of significant responses.

 

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