PBMCs are isolated from human blood and used as effector cells in this assay, which measures the ability of antibodies to induce cell-mediated lysis of target cells. Our service employs advanced techniques to ensure high sensitivity and specificity, allowing for accurate characterization of an ...
Three considerations affected our new assay design: 1) normal B cells within PBMCs introduce 'cold target' competition of radiolabeled tumor B cells if anti-B cell antibodies are present in the ADCC assays; 2) type 1 antibodies to CD20 B cell epitopes are internalized and cleared from the ...
NK effector cells were isolated from PBMCs using the Human NK Cell Isolation (negative selection) Kit 130–092-657 (Miltenyi Biotech, San Diego, CA), according to the manufacturer’s protocol. Median NK cell yield after isolation was typically 0.5-1 × 107 with 94–98% viability. ...
For bioluminescence-based assay by luciferase, 50,000 CEM CD4+ target (T) cells plated in a 96-well V-bottom plate were exposed to NAbs (up to 50 μg/ml) or human control IgG Ab for 30 min at room temperature and then incubated with PBMCs (effectors) at a ratio of 10 ...
PBMCs were used in a conventional cytolytic assay (at different E:T ratios) against the three BC cell lines SKBR3, MCF-7 and BT474, expressing different levels of reactivity with trastuzumab, as shown by a titration assay using this mAb at different concentrations (from 2 to 2 × 10−...
R (45) and ADCC assays with natural killer (NK) cells and peripheral blood monocytes (PBMC) of Fc?RIIIA genotyped donors have been described previously (12). The assay used to measure binding to human C1q has also been previously described (46). Immunofluorescence staining of purified NK ...
When donor PBMC were exposed to TriKE, only NK cells, but not T-cells showed a proliferation-specific pattern in a flow-cytometric assay (Figure 8a). Four additional donors were tested identically to determine whether these findings were reproducible, see Supplementary Figure S1a–d. The results...
Coated ADCC assay ADCC activity was detected by the previously described ADCC-GranToxiLux (GTL) assay using antigen-coated cells (Pollara et al., 2011). Whole PBMCs from a healthy donor were used as effector cells for which the FcγRIIIa receptor was genotyped as being homozygous for valin...
Therefore, this protocol describes methods to perform an in vitro ADCC functional assay with iRBCs. Specifically, it includes protocols on how to grow and culture iRBCs, how to culture peripheral blood mononuclear cells (PMBCs), and how to do in vitro PBMC assays to measure NK degranulation ...
Competitive ELISA assay was used to evaluate the ability of ATG-034-S3 to block the interaction of LILRB4 with its ligand ApoE. ADCC assay was PBMC-mediated cytotoxicity and measured by FACS analysis. ADCP assay was M2 macrophage-mediated phagocytosis and measured using FACS analysis. T cell ...