PD-1 Pathway Recombinant Protein Collection

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Pathway Recombinant Protein Collection www.acrobiosystems.com

Content I. Introduction II. Biotinylated Pathway Proteins IIa. ELISA IIb. FAC Sorting IIc. Biopanning III. HPLC-verified Proteins IV. Full-length Protein V. FITC-labeled VI. Full Product List PD-L1 B7-H2 PD-L2 CTLA-4 B7-1 B7-H5

Introduction Immune checkpoint pathway is a focal point of today s cancer research. is one of the best characterized checkpoint proteins. The binding between 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 pathway may help re-engage the immune system to fight against cancer. In recent years, multiple lines of anti- 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 related products for researchers to investigate and target pathway. 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. ELISA ACROBiosystems has developed a range of prebiotinylated 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. Immobilized human (Cat. No. PD1-H5257) at 2 μg/ml (100 μl/well) can bind to biotinylated human PD-L1, & tag (Cat. No. PD1-H82F3) with a linear range of 0.01-0.25 μg/ml. Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-H82F2 Biotinylated Avi PD1-M82F4 Biotinylated Avi >92% SDS 25ug, 200ug PD-L1 PD1-H82F3 Biotinylated PD-L1 Avi PD-L1 PD1-M82F5 Biotinylated PD-L1 Avi

Suggested Pairs Product Desc. Cat. No. Species Structure Pair 1 Recommended for: Coat PD-L1 Detect PD-L1 / B7-H1 Protein, Tag, low endotoxin Biotinylated / PDCD1, Tag & Tag, Avi Tag (AviTag TM ) PD1-H5258 PD1-H82F2 Avi Pair 2 Recommended for: Coat PD-L1 Detect PD-L1 / B7-H1 Protein, Tag PD1-M5251 Biotinylated / PDCD1, Tag, Avi Tag (AviTag TM ) PD1-M82F4 Avi Pair 3 Recommended for: Coat PD-L1 Detect / PDCD1 Protein, Tag (HPLCverified), low endotoxin Biotinylated PD-L1 / B7-H1, Tag & Tag, Avi Tag (AviTag TM ) PD1-H5257 PD1-H82F3 Avi Pair 4 Recommended for: Coat PD-L1 Detect / PDCD1 Protein, Tag PD1-M5259 Biotinylated PD-L1 / B7-H1, Tag, Avi Tag (AviTag TM ) PD1-M82F5 Avi To further simplify your research, we ve developed a [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 protein. This product is uniquely suitable for rapid and high-throughput screening of putative and PD-L1 inhibitors. Principle of [Biotin]: PD-L1 Inhibitor Screening Kit

Molecule Cat. No. Product Desc. Species Size & PD-L1 EP-101 [Biotin]: PD-L1 Inhibitor Screening Pair 96 tests, 480 tests Immobilized PD-L1 at 2 μg/ml (100 μl/well) can bind biotinylated human with a linear range of 0.038-0.6 μg/ml when detected by Streptavidin-HRP. Inhibition of -PD-L1 binding by an anti- neutralizing antibody is measured by the [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 newly published US patent titled "ANTI- 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 expressing cells. This assay provides measurement for the binding activity of the corresponding antibodies. Biotinylated PD-L1 and PD-L2 in FAC sorting

Biopanning Biopanning has gradually become the mainstream approach for antibody development. The AviTag TM 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 tag is generally undesired for these experiments because it generates false positive results. We have developed AviTag TM pathway proteins with only tag attached. These products are recommended specifically for biopanning. Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-H82E4 Biotinylated Avi PD-L1 PD1-H82E5 Biotinylated PD-L1 Avi PD-L2 PD2-H82E8 Biotinylated PD-L2 Avi Biopanning Workflow with Biotinylated Proteins

HPLC-verified Proteins The quality of recombinant proteins is essential in pharmaceutical assay development. In many situations, SDS-PAGE alone is not sufficient for quality control purposes. To address this need, ACROBiosystems has developed a collection of HPLC-verified and PD-L1 proteins. The purity of PD-L1 Protein, mouse IgG1 Tag, low endotoxin (Cat. No. PD1-H52A3) was more than 90% as determined by SEC-HPLC. Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-H5221 >95% SDS PD1-H5257 >95% SDS PD-L1 PD1-H5229 PD-L1 >98% SDS PD-L1 PD1-H52A3 PD-L1 m >92% SDS PD-L2 PD2-H5251 PD-L2 >95% SDS

Full-length Protein ACROBiosystems newly added the world's first full-length human protein. The full-length protein demonstrates a much higher binding affinity against PD-L1 than the ECD protein. Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-HC214 Full-length, Tag free (25-288 ) >92% SDS 20ug, Full Length, Tag Free, ECD, Tag Extracellular domain (25-170) Transmembrane domain (171-191) Intracellular domain (192-288) Tag Protein structures of full-length human (Cat. No. PD1-HC214) and human ECD protein (Cat. No. PD1-H5221). Immobilized full-length human (Cat. No. PD1-HC214, 20 ng/well) can bind human PD-L1 (Cat. No. PD1-H52A3) with a linear range of 1.2-78 ng/ml, which is much higher than the ECD protein (linear range 250 ng-1,250 ng/ml). FITC-labeled Biotinylated can be used with fluorescence-labeled streptavidin in FACS and immunostaining experiments. However, this process involves multiple rounds of incubation and washes, making it timeconsuming. ACROBiosystems offers an FITC-labeled protein, which allows one-step fluorescence detection. Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-HF257 FITC-labeled >95% SDS 25ug,200ug Immobilized PD-L1, tag (Cat. No. PD1-H5258) at 10 μg/ml (100 μl/well) can bind FITC-labeled, tag (Cat. No. PD1-HF257) with a linear range of 0.3-5 μg/ml.

Full Product List Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-H82E4 Biotinylated Avi PD1-H82F2 Biotinylated Avi PD1-HC214, Tag Free, Full Length (25-288 ) >92% SDS 20ug PD1-H5221 >95% SDS PD1-H5257 >95% SDS PD1-H5284 Twin-Strep >95% SDS 25ug, 250ug PD1-HF257 FITC-labeled PD1-M82F4 Biotinylated Avi >92% SDS 25ug, 200ug PD1-M5228 PD1-M5259 >92% SDS PD1-M8259 Biotinylated >92% SDS 25ug, 200ug PD1-M52A4 (21-167) m PD1-R5253 Rat Rat PD1-R52H2 Rat Rat PD1-RB5251 Rabbit Rabbit PD1-R52H0 Rabbit Rabbit

Molecule Cat. No. Product Desc. Species Structure Purity Size PD1-C5223 Cynomolgus Cynomolgus >95% SDS 50ug, 1mg PD1-C5254 Cynomolgus Cynomolgus PD-L1 PD1-H82E5 Biotinylated PD-L1 Avi PD-L1 PD1-H82F3 Biotinylated PD-L1 Avi PD-L1 PD1-H9229 C13 and N15-labeled, PD-L1 >95% SDS 5ug, 20ug PD-L1 PD1-H5229 PD-L1 >98% SDS PD-L1 PD1-H52A3 PD-L1 m >92% SDS PD-L1 PD1-H5258 PD-L1 PD-L1 PD1-H5282 PD-L1 Twin-Strep >95% SDS 25ug, 250ug PD-L1 PD1-M5220 PD-L1 PD-L1 PD1-M82F5 Biotinylated PD-L1 Avi PD-L1 PD1-M5251 PD-L1 >95% SDS 200ug, 1mg PD-L1 PD1-M52A2 PD-L1 m PD-L1 PD1-C5253 Cynomolgus PD-L1 Cynomolgus >90% SDS PD-L1 PD1-C52H4 Cynomolgus PD-L1 Cynomolgus PD-L2 PD2-H5251 PD-L2 >95% SDS

Molecule Cat. No. Product Desc. Species Structure Purity Size PD-L2 PD2-H82E8 Biotinylated PD-L2 Avi PD-L2 PD2-H5220 PD-L2 PD-L2 PD2-H52A5 PD-L2 m PD-L2 PD2-M52A5 PD-L2 m PD-L2 PD2-M5254 PD-L2 / B7-DC PD-L2 PD2-M52E3 PD-L2 & PD-L1 EP-101 [Biotin]: PD-L1 Inhibitor Screening Pair 96tests, 480tests B7-H5 PD-L2 CTLA-4 B7-H2 PD-L1 B7-1

+ 1 800-810-0816 (US/Canada) + 86 400-682-2521 (Asia & Pacific) techsupport@acrobiosystems.com www.acrobiosystems.com 1 Innovation Way, Newark, DE 19711 PD1-EN02E01-OPE-2017-07-19