REVVITY AL356F

The AlphaLISA® human PD-1/PD-L1 binding kit is designed for the detection of binding activity between human PD-1 and PD-L1, using a fast and simple homogeneous AlphaLISA® assay (no wash steps). This assay can be used to screen for small molecules that inhibit binding, as a competitive ligand binding (CLB) assay to screen therapeutic blocking antibodies, and for potency assays.

AlphaLISA® technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In an AlphaLISA® assay, a biotinylated PD-1 binds to the Streptavidin-coated Alpha Donor beads, while His tagged PD-L1 is captured by Anti-His AlphaLISA® Acceptor beads. When PD-L1 binding to PD-1 happens, Donor beads and Acceptor beads come into close proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615nm.

Programmed death ligand 1 (PD-L1), also known as "cluster of differentiation 274" (CD274) or B7 homolog1 (B7-H1) belongs to the growing B7 family of immune proteins and is expressed in tumor cells. PD-L1 binds to its receptor, PD-1, found on activated T cells, B cells, and myeloid cells, to modulate activation or inhibition. By binding to PD-1, PD-L1 may inhibit ongoing T-cell responses by inducing apoptosis and arresting cell-cycle progression and thus contribute to cancer growth. Monoclonal antibodies targeting PD-1 and PD-L1 are being developed as a means to boost the immune system for the treatment of non-small-cell lung cancer, melanoma, bladder, renal, and triple-negative breast cancers.

Features:

• No-wash steps, no separation steps
• Ease-of-use: few addition steps, fast assay development
• Broad range of affinities: detect strong or weak interactions, from pM to mM affinity
• Distance: measure very large protein or antibody complexes; spanning up to 200nm or more
• High avidity: multiple binding sites on each bead enables use of nanomolar concentrations of antibodies or proteins, as well as use of low affinity binders