PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair, 96 tests

Introduction:
Immune checkpoint pathway is a focal point of today’s cancer research. PD-1 is one of the best characterized checkpoint proteins. The binding between PD-1 and its ligand PD-L1 suppresses T-cell activation and allows cancer cells to escape from body’s immune surveillance. Therefore, the pharmaceutical inhibition of PD-1 or its ligand has been considered a promising strategy by many oncologists.

Reconstitution:
See Certificate of Analysis for details of reconstitution instruction and specific concentration.

Storage:
See Certificate of Analysis for details of storage conditions.
Be sure to store each component at the proper tempe rature upon arrival
Avoid freeze/thaw cycles upon reconstituted.

Stability:
No activity loss was observed after storage at:
Room temperature (RT) for 1 month in lyophilized state.  -20°C for 1 year in lyophilized state.  -80°C for 2 months under sterile conditions after reconstitution.

Application:
This pair is useful for screening for inhibitors of human PD-1 binding to human PD-L1.

Description:
This inhibitor screening ELISA pair is designed to facilitate the identification and characterization of new PD-1 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human PD-1 to immobilized human PD-L1 in a functional ELISA assay, and employs a simple colorimetric sandwich ELISA platform. Briefly, we provide you with a human PD-1-Biotin protein, a human PD-L1 protein, an anti-PD-1 neutralizing antibody (as method verified Std.), and streptavidin-HRP reagent. Your experiment will include 4 simple steps: a) Coat the plate with human PD-L1. b) Add your molecule of interest to the tests. c) Add Human PD-1-Biotin to bind the coated human PD-L1. d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate. Finally, the ability of your compound to inhibit PD-1 : PD-L1 binding will be determined by comparing OD readings among different experimental groups.

Quality Assurance:
Binding of Biotinylated Human PD-1 to Immobilized Human PD-L1 in a Functional ELISA AssayImmobilized human PD-L1 protein at 2 μg/mL (100 μL/well) can bind human PD-1-Biotin with a linear range of 0.038 - 0.6 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting.Inhibition of PD-1-PD-L1 binding by Anti-PD-1 Neutralizing Antibody measured using the PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair (Catalog # EP-101) Serial dilutions of anti-PD-1 neutralizing antibody (1:2 serial dilutions, from 10 μg/mL to 0.078 μg/mL (66.667 nM to 0.521 nM) was added into PD-L1 : PD-1-Biotin binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points prior to log transformation and curve fitting.