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HTRF Human PD1/PD-L1 Binding Kit, 500 Assay Points

This immune checkpoint assay is designed to identify human PD1/PD-L1 checkpoint inhibitors.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Product Variants
Part number: 64PD1PEG
Unit Size: 500 Assay Points
List price: USD 956.69
Your price:
USD 956.69
USD 956.69 /each
Part number: 64PD1PEH
Unit Size: 10,000 Assay Points
List price: USD 12,700.00
Your price:
USD 0.00
USD 12,700.00 /each

Overview

Programmed cell death protein 1 (PD1) is an immune checkpoint receptor that regulates T cell response. Its ligand, programmed death-ligand 1 (PD-L1), is commonly over-expressed on a tumor cell surface. When PD1 is bound to PD-L1, T cell response is suppressed, contributing to tumor immune resistance. Checkpoint inhibitors blocking PD1/PD-L1 complex formation are generating considerable interest in cancer immunotherapy.

Specifications

Assay Points
500
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Shipping Conditions
Shipped in Dry Ice
Therapeutic Area
Oncology & Inflammation
Unit Size
500 Assay Points

Video gallery

How it works

Assay principle

The PD1/PD-L1 binding assay includes tagged human recombinant immune checkpoint partners (PD1 and PD-L1) and labelled anti-tag reagents for HTRF detection. Without an inhibitor, PD1 binds to PD-L1, and the binding of each detection reagent to its tagged target generates an HTRF signal. In the presence of compound, standard, or an antibody blocking the PD1 /PD-L1 interaction, the HTRF signal decreases.

assay-principle-human-pd1-pdl1-binding-kit-64spd1peg

 

Assay protocol

This PD1/PD-L1 binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate, and the tagged PD1 & PD-L1 proteins are then added, followed by the dispensing of the HTRF reagents. The reagents labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.

assay-protocol-human-pd1-pdl1-binding-kit-64spd1peg

 

Assay validation

Inhibitory effect of blocking antibodies and small molecules

The inhibitory effects of three antibodies and two small molecules known to inhibit the PD1 / PD-L1 interaction were tested. The two anti-PD1 antibodies (pembrolizumab & nivolumab) were shown to inhibit the interaction with IC50 values of 2.0 & 2.4 nM respectively. The anti-PD-L1 antibody (atezolizumab) disrupted the interaction, with an IC50 of 3.9 nM. Two small molecule PD1 / PD-L1 inhibitors 1 & 3 show IC50s of 177 & 146 nM in these assay conditions.

assay-validation-human-pd1-pdl1-binding-kit-1

 

Inhibitory effect of the PD1 / PD-L1 standard

The PD1 / PD-L1 standard stock solution provided in the kit was diluted to prepare the 8-point dose response curve using 5-fold serial dilutions. As described in the assay protocol, standards were dispensed into the assay plate, and the tagged PD1 & PD-L1 proteins were then added, followed by the dispensing of the HTRF reagents. The data presented in the graph show that the PD1 / PD-L1 interaction is inhibited, with an IC50 of 3.8 nM.

assay-validation-human-pd1-pdl1-binding-kit-2

 

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SDS, COAs, Manuals and more

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