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

For research use only.
Materials Provided
IDComponentsSize (96 tests)Size (480 tests)
A001-214Human PD-L125 μg125 μg
A002-214Human PD-1-Biotin5 μg25 μg
A003-214Streptavidin-HRP10 μg25 μg
PD1-NA001Anti-PD-1 Neutralizing Antibody20 μg100 μg
  • Background
    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.
  • Application

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

  • 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 temperature upon arrival.

    It is recommended not to freeze thaw more than 3 times.

    This product is stable after storage at:

    • Room temperature (RT) for 1 month in lyophilized state;

    • -20℃ for 1 year in lyophilized state;

    • -70℃ for 6 months under sterile conditions after reconstitution.

  • Assay Principles

    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 Human PD-1-Biotin to bind the coated human PD-L1.

    • c) Add your molecule of interest to the tests.

    • 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.

Typical Data

Binding of Biotinylated Human PD-1 to Immobilized Human PD-L1 in a Functional ELISA Assay

Immobilized 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 (Cat. No. PD1-NA001) measured using the PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair (Cat. No. EP-101)

Serial dilutions of anti-PD-1 neutralizing antibody (Cat. No. PD1-NA001) (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.

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