T-cell based immunotherapy strategies have become increasingly important in disease treatments that harness the human immune system. LakePharma offers a variety of assays for characterizing T-cell activity with candidate antibodies.
1. T-cell activation and proliferation assays include stimulation with staphylococcal enterotoxin B (SEB), cytomegalovirus (CMV) recall antigen, and anti-CD3 antibodies.
2. Assay readouts include cytokine measurement by ELISA or FACS.
3. Mixed lymphocyte reaction (MLR) and T-regulatory suppression assays.
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View LakePharma T-Cell Functional Assay Case Studies.
T-CELL STIMULATION ASSAY
1. PBMCs, SEB, and test antibodies are co-incubated for 3 days
2. IL-2 concentrations measured by ELISA or FACS
T-CELL PROLIFERATION ASSAY
1. PBMCs, anti-CD3 antibodies (can be plate bound, soluble, or anti-CD3 coated beads), test antibodies, and CSFE dye are co-incubated for 3 days
2. Samples harvest and stained with anti-CD4
3. T-cell proliferation assessed by FACS
CMV RECALL ASSAY
1. Incubate PBMCs from CMV-positive donors with CMV pp65 peptide pool and test antibodies for 3 days.
2. IFN-γ ELISA performed on collected supernatants.
MIXED LYMPHOCYTE REACTION
1. Macrophage and dendritic cell generation from two individual donors and FACS validation.
2. Coculture at single ratio with allogeneic frozen PBMCs from 2 individual donors (4 pairwise combinations + controls)
3. Add test and control antibodies.
4. IL-2 ELISA performed
T-REG SUPPRESSION ASSAY
1. PBMCs isolated, T-reg population sorted.
2. Unsorted PBMCs labeled with CFSE
3. Co-incubation with T-reg cells, PBMCs, test antibodies for 4 days
4. T-reg suppression measured by FACS
LakePharma T-Cell Functional Assay Case Studies