human ifn γ elisa  (R&D Systems)

Journal: Biomolecules

Article Title: High-Affinity Fully Human Anti-EpCAM Antibody with Biased IL-2 Exhibits Potent Antitumor Activity

doi: 10.3390/biom14111399

Figure Lengend Snippet: Anti-EpCAM-IL-2v immunocytokines show potent activation of immune cells in vitro. ( A , B ) Binding kinetics of m801, m801.2, and m801.3 to the immobilized IL-2Rβ ( A ) or IL-2Rα ( B ) as determined by biolayer interferometry. The antibody concentrations used were 100 nM (orange), 33.3 nM (blue), 11.1 nM (purple), and 3.7 nM (green). ( C ) Flow cytometry analysis of CD69 expression in effector cells treated with varying concentrations of m801, m801.2, and m801.3. Isotype control: mAb CR3022. ( D ) IFN-γ secretion levels in PBMC culture supernatants following stimulation with varying concentrations of anti-EpCAM-IL-2v immunocytokines or control antibodies. Each data point is shown as mean ± SD of three biological replicates.

Article Snippet: In brief, PBMC were incubated with increasing concentration of antibodies for 72 h. Supernatant from the assay plates was collected to determine the IFN-γ concentration by IFN-γ ELISA test kit (#DIF50C, R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Activation Assay, In Vitro, Binding Assay, Flow Cytometry, Expressing, Control