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.
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This kit is designed for the detection of binding activity between PD-1 and PD-L1 using a homogeneous AlphaLISA assay (no wash steps). This assay can facilitate the design and development of therapetics by using competitive binding.
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 615 nm.
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.
|Assay Target||PD-1, PD-L1|
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||500 assay points|
Mouse pharmacological models continue to play a large role in the study of human disease, and mouse tool reagents have shown high utility in immunology and cancer research for decades. It can often be quicker to learn about immunology and the regulation of immune responses using a syngeneic mouse model. However, working in mouse systems can often require the development of separate mouse reagents, if the therapeutic agent of interest does not cross-react with mouse. Find out how the AlphaLISA® human PD-1/PD-L1 and AlphaLISA mouse PD-1/PD-L1 binding assays provide a fast, powerful, homogeneous platform for obtain binding potencies from potential novel drug candidates.
Many tumor cells, which under normal, healthy conditions would be recognized by the body’s T cells and thereby targeted for destruction, have developed ways to evade the host immune system by taking advantage of immune checkpoint pathways. Among the most promising approaches to activating therapeutic antitumor immunity is through the blockade of immune checkpoints. The programmed cell death-1 (PD-1) immune checkpoint pathway is a negative regulator of T-cell immune function. When PD-1 is bound to programmed cell death-ligand 1 (PD-L1), T cell response is suppressed. Inhibitors that block PD-1/PD-L1 complex formation lead to increased activation of T-cells and immune system functions, allowing the body’s immune system to identify and attack tumor cells. So far, several anti-PD-1 or PD-L1 monoclonal antibodies have been developed to treat a variety of cancers, including non-small cell lung carcinoma (NSCLC), metastatic melanoma and renal cancer. The promise of therapeutically exploiting this pathway has created a need for more robust, straight-forward assays to identify and qualify potential inhibitors which interrupt PD-1/PD-L1 binding. Find out how AlphaLISA® technology provides a simple, homogenous, straightforward method for detecting PD-1/PD-L1 binding.
Immune checkpoints serve a critical role in the immune system to prevent autoimmunity and manage the degree and duration of an immune response. Cytotoxic T-Lymphocyte-associated protein 4 (CTLA-4 or CD152) is an inhibitory transmembrane protein involved in an immune checkpoint of significant interest for therapeutic development. When CTLA-4 is expressed and competes with CD28, the immune system response is downregulated. As a result of this immune system response balance, immune checkpoints provide an opportunity for therapeutic intervention to modulate immune system activity.
There is a high demand for new drugs to block CTLA-4 and modulate immune system activity. In this application note, we demonstrate how to screen for novel CTLA-4 blocking drugs by utilizing the AlphaLISA CTLA-4/CD80 binding assay.