PD-1/PD-L1 immune checkpoint Bioassay
Targeting immune checkpoint receptors have shown great promise in the management of immune-mediated effects on cancer cells. These receptors play a key regulatory role in maintaining immune equilibrium. Recently immunotherapies aimed at blocking immune checkpoints PD-1 and CTLA-4 have demonstrated success in the management of various cancers.
Our team has the capabilities to offer suitable robust and consistent immune cell line-based bioassays to our clients to screen the potency of their biologics. These can be further validated in a primary cell model.
The PD-1/PD-L1 Blockade Bioassay is a biologically relevant MOA-based assay that can be used to measure the potency and stability of antibodies and other biologics designed to block the PD-1/PD-L1 interaction.
For more information on how Cellomatics can support your PD-1/PD-L1 bioassay project, click the link below.
PD-1/PD-L1 immune checkpoint bioassay
The PD-1/PD-L1 luciferase bioassay enables detection of the inhibitory activity of anti-PD-1 and anti-PD-L1 blocking antibodies as quantified by increases in luminescence. In the assay workflow, PD-L1 expressing aAPC/CHO-K1 cells target cells are plated and treated with increasing concentrations of test antibodies along with the PD-1 expressing effector cells. Luminescence is detected after 6 hours using Bio-Glo™ Reagent and the GloMax® Discover System. Validation of the assay has been performed using Nivolumab and Pembrolizumab. EC50 for Nivolumab and Pembrolizumab is 0.28 µg/ml and 0.11 µg/ml respectively (a). The assay is specific for PD-1 blockade as confirmed by incubation with Ipilumamb, a CTLA-4 inhibitory antibody. A high degree of intra-plate reproducibility in Nivolumab inhibitory response is demonstrated (b).
PBMCs proliferate in response to anti-CD3 stimulation
PBMCs were treated for 96 hours with increasing concentrations of anti-CD3 antibody (0 µM, 0.25 µM, 1 µM and 5 µM; black bar) or with Isotype control (blue bar) or with Nivolumab (red bar). Supernatants were collected and IFN-γ release was quantified by ELISA. Results demonstrate a dose dependent increase in IFNγ levels which are further potentiated by PD-1 blockade as suggested by Nivolumab treatment.
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Our experienced team of in vitro laboratory scientists will work with you to understand your project and provide a bespoke project plan with a professional, flexible service and a fast turnaround time.
To request a consultation where we can discuss your exact requirements, please contact Cellomatics Biosciences.