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PD-1 Pathway Recombinant Protein Collection

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 and/or PD-L2 suppresses T-cell activation, and allows cancer cells to escape from body’s immune surveillance. This mechanism prompts the idea that neutralizing PD-1 pathway may help re-engage the immune system to fight against cancer.

In recent years, multiple lines of anti-PD-1 drugs have been developed, and have demonstrated enormous potential in recent clinical trials. Patients with melanoma, renal cell carcinoma, non-small cell lung cancer (NSCLC), or hematological cancers all respond positively to this treatment.

ACROBiosystems provides a comprehensive series of PD-1 related products for researchers to investigate and target PD-1 pathway.

PD1 Pathway Targets

Key Features

Biotinylated PD-1 Pathway Proteins

ELISA

ELISA is an assay technique routinely used to characterize protein-protein interaction. It’s often used in initial high-throughput screening (HTS) of compound library. ACROBiosystems has developed a range of pre-biotinylated PD-1 pathway proteins. Biotinylated proteins can serve as two-way antibodies for both capture and detection in ELISA assays. They demonstrate higher specificity and detection sensitivity than traditional antibodies.

PD1-H82F3-E1.jpg

Fig. 1 Immobilized human PD-1 (Cat. No. PD1-H5257) at 2 μg/mL (100 μL/well) can bind to biotinylated human PD-L1, His & Fc tag (Cat. No. PD1-H82F3) with a linear range of 0.01-0.25 μg/ml.



Suggested Pairs

Product Description Cat. No. Structure
Pair 1 Human PD-L1 / B7-H1 Protein, Fc Tag, low endotoxin PD1-H5258 PD1-H5258-STC1
Pair 2 Mouse PD-L1 / B7-H1 Protein, Fc Tag PD1-M5251 PD1-M5251-STC1
Biotinylated Mouse PD-1 / PDCD1, Fc Tag, Avi Tag (AviTagTM) PD1-M82F4 PD1-M82F4-STC1
Pair 3 Human PD-1 / PDCD1 Protein, Fc Tag (HPLC-verified), low endotoxin PD1-H5257 PD1-H5257
Biotinylated Human PD-L1 / B7-H1, His Tag & Fc Tag, Avi Tag (AviTagTM) PD1-H82F3 PD1-H82F3-STC1
Pair 4 Mouse PD-1 / PDCD1 Protein, Fc Tag PD1-M5259 PD1-M5259
Biotinylated Mouse PD-L1 / B7-H1, Fc Tag, Avi Tag (AviTagTM) PD1-M82F5 PD1-M82F5-STC1

To further simplify your research, we’ve developed a PD-1[Biotin]: PD-L1 Inhibitor Screening Kit (Cat. No. EP-101). This assay employs a simple colorimetric ELISA platform, which measures the binding between immobilized human PD-L1 and in-house developed biotinylated PD-1 protein. This product is uniquely suitable for rapid and high-throughput screening of putative PD-1 and PD-L1 inhibitors.

Molecule Cat. No. Product Description Size
PD-1 & PD-L1 EP-101 PD-1[Biotin]: PD-L1 Inhibitor Screening Pair 96 tests, 480 tests
PD-L1

Fig. 2 Immobilized PD-L1 at 2 μg/mL (100 μL/well) can bind biotinylated human PD-1 with a linear range of 0.038 - 0.6 μg/mL when detected by Streptavidin-HRP.


PD-1-PD-L1.jpg

Fig. 3 Inhibition of PD-1-PD-L1 binding by an anti-PD-1 neutralizing antibody is measured by the PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair (Cat. No. EP-101).


FAC Sorting

Biotinylated proteins can be used along with fluorophore-tagged SA to detect/isolate cells expressing particular surface markers. In a US patent titled "ANTI-PD-1 ANTIBODIES AND METHODS OF USE THEREOF" (US patent 20160159905), the authors (from Pfizer Inc.) used ACROBiosystems' biotinylated PD-L1 (Cat. No. PD1-H82F3) and biotinylated PD-L2 (Cat. No. PD2-H82F6) to compete with candidate antibodies in a binding assay against PD-1 expressing cells. This assay provides measurement for the binding activity of the corresponding antibodies.

Biopanning

Biopanning has gradually become the mainstream approach for antibody development. The AviTagTM biotinylated proteins can be used with SA-coated magnetic beads/surface in biopanning. It’s a much better choice than non-biotinylated, or chemically biotinylated proteins because it offers higher coating density and uniformed antigen presentation.

Please note that the Fc tag is generally undesired for these experiments because it generates false positive results. We have developed AviTagTM PD-1 pathway proteins with only His tag attached. These products are recommended specifically for biopanning.

Molecule Cat. No. Structure
PD-1 PD1-H82E4 PD1-H82E4-STC1
PD-L1 PD1-H82E5 PD1-H82E5-STC1
PD-L2 PD2-H82E8 PD2-H82E8-STC1

Cell based assay verified proteins

As a professional supplier of pharmaceutical companies, we have established a cell based assay platform to verify the activity of the target protein, which provide cell based experiment data support for drug screening.

ACROBiosystems used human PD-1 overexpressed 293 cells made in-house in flow cytometry assay and cell based ELISA, and the results showed good performance of PD-L1 (Cat. No. PD1-H5258) and PD-L2 (Cat. No. PD2-H5251) binding to PD-1.

FACS Analysis of PD-L1 or PD-L2 binding to cell surface PD-1
PD1-H5258

Fig. 4 Flow Cytometry assay shows that recombinant human PD-L1 (Cat. No. PD1-H5258) can bind to 293 cell overexpressing human PD-1.The concentration of PD-L1 used is 10ug/mL


PD2-H5251

Fig. 5 Immobilized cell surface PD-1 (5x104 of cells per well) can bind Human PD-L2 Protein, Fc Tag (Cat. No. PD2-H5251) with an EC50 of 0.018 μg/mL (Routinely tested).


Validated applicable for screening of neutralizing antibody
PD1-H82F3-E1.jpg

Fig. 6 FACS analysis shows that the binding of PD-L1 (Cat. No. PD1-H5258) to 293 overexpressing PD-1 was inhibited by increasing concentration of neutralizing anti-hPD-L1 antibody. The concentration of PD-L1 used is 10 μg/mL. The IC50 is 12.92 μg/mL.

Cell based ELISA
PD1-H5258

Fig. 7 Immobilized cell surface PD-1 (5x104 of cells per well) can bind Human PD-L1, Fc Tag (Cat. No. PD1-H5258) with an EC50 of 0.029 μg/mL.


PD2-H5251

Fig. 8 Immobilized cell surface PD-1 (5x104 of cells per well) can bind Human PD-L2, Fc Tag (Cat. No. PD2-H5251) with an EC50 of 0.018 μg/mL.


latest citations

References

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FAQ

Q

What research applications can PD-1, PD-L1, and PD-L2 recombinant proteins support?

Recombinant PD-1, PD-L1, and PD-L2 proteins can support receptor–ligand binding studies, antibody characterization, competition or inhibition assays, and related assay development. ACROBiosystems provides these pathway proteins in multiple species, tag, and labeling formats for different experimental configurations. Reagent selection should begin with the PD-1–PD-L1 or PD-1–PD-L2 interaction being investigated and the intended assay readout.
Q

How can cross-species binding and ligand-blocking activity of anti-PD-1 antibodies be characterized?

Cross-species binding can be assessed using ELISA, SPR, or BLI, while PD-L1 and PD-L2 blocking activities should be evaluated separately. ACROBiosystems provides bioactivity-validated PD-1 proteins across multiple species, tags, and labeling formats to support antibody cross-reactivity and affinity profiling. Its human PD-1/PD-L1 and PD-1/PD-L2 TR-FRET kits enable rapid, wash-free inhibition assays within 1 hour, with reference antibody validation supporting reliable ligand-blocking comparisons.
Q

How should I choose protein formats for PD-1–PD-L1 or PD-1–PD-L2 binding assays?

Protein format should be matched to the planned immobilization, capture, and detection strategy. ACROBiosystems provides unconjugated and biotinylated pathway proteins in different tag configurations, enabling assay designs such as immobilized target proteins paired with biotinylated interaction partners for streptavidin-based detection. Researchers should review product-specific binding data because proteins with different constructs or labeling formats are not necessarily validated for the same assay configuration.
Q

What is needed to assess FcγR involvement in PD-1 agonist antibody activity?

PD-1 agonist activity can depend on FcγR-mediated crosslinking and should be evaluated through FcγR binding and functional T-cell assays. ACROBiosystems offers SPR/BLI-validated FcγRI (CD64), FcγRII (CD32a/b), and FcγRIII (CD16) proteins for comparative Fc-binding profiling, alongside CD32b-expressing stable cells for crosslinking studies. Comparing functional responses with and without FcγR engagement helps establish its contribution to PD-1 agonism.
Q

How can biochemical and cell-based assays complement each other when evaluating PD-1 pathway interactions?

Biochemical assays can characterize PD-1 binding to PD-L1 or PD-L2 under defined conditions, while cell-based assays can evaluate binding or inhibition when the interaction partner is presented on a cell surface. Selected ACROBiosystems PD-L1 and PD-L2 products include product-specific data using PD-1-expressing cells in addition to biochemical binding or competition assays. Validation in one assay format should not be assumed to establish performance in another.
Q

How can anti-PD-L1 antibodies be characterized for blockade of both PD-1 and CD80 interactions?

PD-L1 interacts with both PD-1 and CD80, but inhibition of one interaction does not establish blockade of the other. Both interactions should be assessed independently. ACROBiosystems provides bioactivity-validated PD-L1 and CD80 proteins in multiple formats, including biotinylated proteins for competition assays, alongside PD-1/PD-L1 inhibitor screening kits. These reagents support comparative antibody blocking profiles, while cell-based assays can further assess disruption of the physiological PD-L1–CD80 cis interaction.
  • Introduction
  • PD1 Pathway Targets
  • Key Features
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