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Inhibitor Screening Kits

Background

In some immune checkpoint pathway, tumor cells can hijack the checkpoint pathways to avoid attacks from the immune system. For example, 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.

Not only in the immune checkpoint proteins, but also the protein such as PCSK9, which binds to the epidermal growth factor-like repeat A (EGF-A) domain of the low-density lipoprotein receptor (LDLR), inducing LDLR degradation. Inhibition of PCSK9 protein function is currently being explored as a means of lowering cholesterol levels.

Antibody drugs can produce therapeutic effects by blocking or neutralizing the interaction between proteins. In response to this market demand, ACROBiosystems developed ELISA kits that can screen neutralizing antibodies.

Product List
Cat.No.Product DescriptionSizeAssay range
(μg/mL)
Price
EP-101PD-1[Biotin]: PD-L1 Inhibitor Screening Pair ELISA Assay Pair96 tests/480 tests10-0.078$600/$2150
EP-102CD47: SIRP alpha [Biotinylated] Inhibitor Screening ELISA Assay Pair96 tests/480 tests10-0.04$600/$2150
EP-103PCSK9 [Biotinylated]: LDL R Inhibitor Screening ELISA Assay Pair96 tests/480 tests20-0.04$600/$2150
Assay Principle
The assay employs a simple colorimetric ELISA platform which is based on the competitive binding technique
Take PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair (EP-101) for example.
Your experiment will include 4 simple steps:
  • Coat the plate with human PD-L1.

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

  • Add your molecule of interest to the tests.

  • 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.
Application
EP-101 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 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 (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.

Features
Fast and Easy-to-Use
By employing the biotin pre-labeled proteins, we eliminate the often time-consuming labeling process, and greatly simplify the experimental procedures.
Authentic Human Protein Interaction
Proteins in the three inhibitor screening kits are expressed in the HEK293 cells. The use of human cells as expression host confers authentic post-translational modifications essential for their binding activities.
Neutralizing Antibody Included
Each inhibitor screening kit includes the neutralizing mAb as a standard positive control. This would allow the users to validate their experiments.
High Throughput and Multiple Sizes Available
We offer both 96 tests and 480 tests as standard package sizes, which enables simultaneous analyses of a large amount of inhibitors.
High Detection Sensitivity and Reproducibility
Our biotin labeling platform produces biotinylated proteins with high detection sensitivity and minimal lot-to-lot variation. These features ensure that the assay kit has desired detection sensitivity and the results are reproducible.

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