PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Kit, 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:
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.

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.

Clinical and Translational Updates

(1) "New advances in the study of PD-1/PD-L1 inhibitors-induced liver injury"
Yue, Li, Li
Int Immunopharmacol (2024) 131, 111799
(2) "Metabolism/Immunity Dual-Regulation Thermogels Potentiating Immunotherapy of Glioblastoma Through Lactate-Excretion Inhibition and PD-1/PD-L1 Blockade"
Li, Xu, Ruan et al
Adv Sci (Weinh) (2024)
(3) "Evaluation of the efficacy and safety of a precise thymalfasin-regulated PRaG regimen for advanced refractory solid tumours: protocol for the open-label, prospective, multicentre study (PRaG5.0 study)"
Kong, Chen, Xu et al
BMJ Open (2024) 14 (3), e075642
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