Antigen-presenting cells (APCs), typically dendritic cells (DCs), can process antigens and present peptide epitopes in conjugation with human leukocyte antigen (HLA) class II molecules to specific naïve helper-T (Th) cells, which results in the activation of Th cells. The activated Th cells can stimulate B cells to produce antibodies against antigens. For instance, anti-drug antibodies (ADAs) are generated in this way. Therefore, detection of T cell responses is frequently employed to monitor the ability of biotherapeutic candidates to elicit immune system activation, i.e., the immunogenicity. The most widely used method is the ex vivo DC-T cell proliferation assay.
Creative Biolabs provides SIAT® ex vivo DC-T cell proliferation assay service to assess the immunogenicity of biotherapeutic drug candidates using cells from HLA-typed donors. The SIAT® DC-T cell proliferation assay allows the use of whole proteins, and more importantly, it can assess the factors other than protein sequence that contribute to immunogenicity. High-resolution HLA II-typed donor blood samples are used to mimic the allele frequencies found in the global population or to provide representative coverage of a particular population of interest without the need of animal models. Information provided by this assay is very useful during the early development of biotherapeutic drugs in terms of lead selection, humanization, and/or de-immunization.
Proliferation and differentiation of naïve T cells upon stimulation by DCs.
(Idris-Khodja et al. 2014)
SIAT® DC-T cell proliferation assay uses Flow Cytometry to detect T cell proliferation after co-culture with loaded DCs, which is extremely sensitive and accurate. Specifically, T cells are labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) dye. Those that proliferate in response to peptide antigen presented by DCs show a CFSE dye dilution, which can be measured directly by Flow Cytometry. In combination with detection of cell surface markers, the percentage of proliferating CD4+ cells, as well as the detailed phenotype of T cell responses are all monitored. Below is a brief description of our workflow for SIAT® DC-T cell proliferation assay service.
With years of experience in biotherapeutic drug development, scientists of Creative Biolabs are confident in offering the best service to meet our clients’ specific requirements.
More SIAT® Immunogenicity Related Services at Creative Biolabs
In Silico Immunogenicity Assessment
In Vitro Class II HLA Binding Assay
Ex Vivo Immunogenicity Assessment
DC-T Cell Proliferation Assay
Antigen Presentation Assay
In Vivo Immunogenicity Assessment
Anti-drug Antibodies (ADA) Assays