PD-1/PD-L1 Blockade Bioassay

J1250_PD-1-3aPD-L1-Blockade-Bioassay_3
J1252
J1250_PD-1-3aPD-L1-Blockade-Bioassay_3
View information on global supply logistics
customize-this-small

Immune Checkpoint Bioassays for Characterizing Immune-Modulating Antibodies

  • Prequalified according to ICH guidelines
  • 96- and 384-well plate formats
  • No primary cell culture required
  • Supplied in convenient thaw-and-use format; also available in cell propagation model (CPM) format

Choose a Product

Size

Catalog number selected: J1250

$ 1,420.00
Your price:
Add to Cart
This product is discontinued
This product is available under our Early Access program - Learn More
This product is available under our Catalog (FT) program - Learn More
PD-1/PD-L1 Blockade Bioassay
Thaw and Use Kits/1X each component
$ 1,420.00
Your price: Log in
Change Configuration

Measure the ability of biologics to block immune checkpoint signals

PD-1 is an immune inhibitory receptor expressed on activated T cells and B cells and plays a critical role in regulating immune responses to tumor antigens and autoantigens. Engagement of PD-1 by either of its ligands, PD-L1 or PD-L2, on an adjacent cell inhibits TCR signaling and TCR-mediated proliferation, transcriptional activation and cytokine production. Therapeutic antibodies and Fc fusion proteins designed to block the PD-1/PD-L1 interaction show promising results in clinical trials for the treatment of a variety of cancers.

The PD-1/PD-L1 Blockade Bioassay is a biologically relevant MOA-based assay that can be used to measure the potency and stability of antibodies and other biologics designed to block the PD-1/PD-L1 interaction.

Control Ab, Anti-PD-1, is a mouse anti-human PD-1 antibody that shows blocking activity for PD-1 with its ligands PD-L1 and PD-L2. It is intended for use as a control with the PD-1/PD-L1 Blockade Bioassay.

aAPC/CHO-K1 Cells are CHO-K1 cells with an engineered cell surface protein designed to activate cognate TCRs in an antigen-independent manner. The PD-L1 Negative aAPC/CHO-K1 Cells can serve as a negative control in the PD-1/PD-L1 Blockade Bioassay.

Assay Components

The thaw-and-use format of the assay (Cat. No. J1250, J1255) offers convenience and minimizes variability. The assay consists of two cell lines:

PD-1 Effector Cells: Jurkat T cells stably expressing human PD-1 and NFAT-induced luciferase.

PD-L1 aAPC/CHO-K1 Cells: CHO-K1 cells stably expressing human PD-L1 and a cell surface protein designed to activate cognate TCRs in an antigen-independent manner.

When the two cell types are co-cultured, the PD-1/PD-L1 interaction inhibits TCR signaling and NFAT-mediated luciferase activity. Addition of an antibody that blocks either PD-1 or PD-L1 releases the inhibitory signal and results in TCR signaling and NFAT-mediated luciferase activity.

The assay is also available in CPM format (Cat. No. J1252) containing cryopreserved cells that can be thawed, propagated and banked for long-term use. 

Workflow

Standard bioassay workflow with cultured cells
Improved bioassay workflow with thaw-and-use cells

Standardized Reagents Improve Results

  • Thaw-and-use format provides significant time and labor savings over traditional methods while reducing variability.
  • Kit includes all of the required reagents in standardized formats.
  • Easily amenable to high-throughput workflows.

Assay Principle

The PD-1/PD-L1 Blockade Bioassay uses an alternative readout from traditional primary cell based assays: the TCR and NFAT-mediated activation of luciferase activity in the effector cells. Primary cells are replaced with a cell line stably expressing TCR, PD-1 and NFAT-induced luciferase.

13342MB-W

Representation of the PD-1/PD-L1 Blockade Bioassay. The bioassay consists of two genetically engineered cell lines, PD-1 Effector Cells and PD-L1 aAPC/CHO-K1 Cells. When co-cultured, the PD-1/PD-L1 interaction inhibits TCR-mediated luminescence. When the PD-1/PD-L1 interaction is disrupted, TCR activation induces luminescence (via activation of the NFAT pathway) that can be detected by addition of Bio-Glo™ Reagent and quantitation with a luminometer.

Assay Performance Data

Accuracy and precision are critical to bioassays and the development of biologics. This assay was developed and prequalified according to ICH guidelines for accuracy and precision, giving it the performance characteristics required for many applications of a bioassay used for antibody drug discovery research and development.

Accuracy: Measure of how close results are to a reference or "target" value.

IntraLab Precision: Measure of repeatability from multiple assays of the same homogeneous sample under the same operating conditions over a short time (same analyst; same lab; same day).

Intermediate Precision: Measure of repeatability of multiple assays of the same homogeneous sample with different analysts or equipment or on different days.

Linearity: Ability of the assay to provide measured values that are proportional to the concentration of the sample.

32454752-data-target-combined

The PD-1/PD-L1 Blockade Bioassay Shows Precision, Accuracy and Linearity.

Parameter Results
Accuracy % Expected Relative Potency % Recovery
50 96.1
75 99.9
150 104.4
200 104.9
Repeatability (% CV) 100% (Reference) 8.5
Intermediate Precision (% CV)   8.7
Linearity (r2)   0.997
Linearity (y = mx + b)   y = 1.088x – 0.084
A 50–200% theoretical potency series of nivolumab (PD-1 blocking antibody) was analyzed in triplicate in three independent experiments performed on three days by two analysts. Bio-Glo™ Reagent was added and luminescence quantified. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells.

 

13070MB-W
The PD-1/PD-L1 Blockade Bioassay demonstrates appropriate rank ordering of antibody potency.  PD-1 Effector Cells and PD-L1 aAPC cells were incubated with anti-PD-L1 antibody preparations representing a 50–200% potency range.
13071MB-W
The PD-1/PD-L1 Blockade Bioassay is reproducible across four independent 96-well plates.  PD-1 Effector Cells and PD-L1 aAPC Cells were incubated with anti-PD-L1 antibody representing 100% potency.

Product Kit Formats

  • PD-1/PD-L1 Blockade Bioassay is available in 1X and 5X sizes.
  • PD-1/PD-L1 Blockade Bioassay is also available in CPM format (Cat.# J1252).
  • Control Ab, Anti-PD-1, is available separately (Cat.# J1201).
  • PD-L1 Negative Cells are available separately (Cat.# J1191, J1195).

Protocols

Complete Protocol

Specifications

You are viewing: J1250 Change Configuration

What's in the box?

Item Part # Size

PD-1 Effector Cells

 J115A 1 × 0.5ml

PD-L1 aAPC/CHO-K1 Cells

 J109A 1 × 0.5ml

Fetal Bovine Serum

 J121A 1 × 4ml

Ham's F-12 Medium

 J123A 1 × 25ml

RPMI 1640 Medium

 G708A 1 × 36ml

Bio-Glo™ Luciferase Assay Buffer

 G719A 1 × 10ml

Bio-Glo™ Luciferase Assay Substrate

 G720A 1 × 1 vial

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. No. 8,008,006.

Patent Pending.

U.S. Pat. No. 10,077,244 and other patents.

Licensed from Lonza Cologne GmbH under U.S. Pat. Nos. 7,700,357, 8,192,990 and 8,003,389, European Pat. Nos. 1297119, 1522587, 1607484 and 1741778 and other pending and issued patents.

Product cannot be used for proficiency testing.

Specifications

You are viewing: J1255 Change Configuration

What's in the box?

Item Part # Size

PD-1 Effector Cells

 J115A 5 × 0.5ml

PD-L1 aAPC/CHO-K1 Cells

 J109A 5 × 0.5ml

Fetal Bovine Serum

 J121A 5 × 4ml

Ham's F-12 Medium

 J123A 5 × 25ml

RPMI 1640 Medium

 G708A 5 × 36ml

Bio-Glo™ Luciferase Assay Buffer

 G719A 5 × 10ml

Bio-Glo™ Luciferase Assay Substrate

 G720A 5 × 1 vial

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

Patent Pending.

U.S. Pat. No. 8,008,006.

U.S. Pat. No. 10,077,244 and other patents.

Licensed from Lonza Cologne GmbH under U.S. Pat. Nos. 7,700,357, 8,192,990 and 8,003,389, European Pat. Nos. 1297119, 1522587, 1607484 and 1741778 and other pending and issued patents.

Product cannot be used for proficiency testing.

Specifications

You are viewing: J1252 Change Configuration

What's in the box?

Item Part # Size

PD-1 Effector Cells (CPM)

 J112A 2 × 1ml

PD-L1 aAPC/CHO-K1 Cells (CPM)

 J108A 2 × 1ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use" means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells except that recipient may propagate and store the cells for long-term use. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega's. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

Patent Pending.

U.S. Pat. No. 8,008,006.

Licensed from Lonza Cologne GmbH under U.S. Pat. Nos. 7,700,357, 8,192,990 and 8,003,389, European Pat. Nos. 1297119, 1522587, 1607484 and 1741778 and other pending and issued patents.

Product cannot be used for proficiency testing.

Specifications

You are viewing: J1191 Change Configuration

What's in the box?

Item Part # Size

aAPC/CHO-K1 Cells

 J119A 1 × 0.5ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

XX

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use; (2) discovery, development and monitoring of biologic drugs and vaccines; (3) quality assurance testing of biologic drugs and vaccines; and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. “Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use luminescent assay reagents purchased from Promega for all luminescence-based assays using this product, or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

Product cannot be used for proficiency testing.

Specifications

You are viewing: J1195 Change Configuration

What's in the box?

Item Part # Size

aAPC/CHO-K1 Cells

 J119A 5 × 0.5ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

XX

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use; (2) discovery, development and monitoring of biologic drugs and vaccines; (3) quality assurance testing of biologic drugs and vaccines; and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. “Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use luminescent assay reagents purchased from Promega for all luminescence-based assays using this product, or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

Product cannot be used for proficiency testing.

Specifications

You are viewing: J1201 Change Configuration

What's in the box?

Item Part # Size

Control Ab, Anti-PD-1

 J120A 1 × 100μg

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

CC

Please see the Protocol for detailed storage conditions.

Resources

Articles

Posters

Other Resources

Similar Products

CTLA-4 Blockade Bioassay

Measure potency and stability of biologics designed to block CTLA-4 interaction with its ligands.

JA3001, JA3005, JA1400, JA1020

LAG-3/MHCII Blockade Bioassay

Measure the potency and stability of biologics designed to block the interaction of LAG-3 and MHCII.

JA1111, JA1115, JA1112, K1150, K1201

TIGIT/CD155 Blockade Bioassay

Measure the potency and stability of biologics designed to block the interaction of TIGIT and CD155.

J2201, J2205, J2092, J2051, J1921

Frequently Used With

GloMax® Discover System

High-performance microplate reader for detecting luminescence, fluorescence and absorbance.

GM3000

Bio-Glo™ Luciferase Assay System

Bioluminescent detection reagent qualified for use with bioassays that utilize firefly luciferase.

G7940, G7941

RealTime-Glo™ MT Cell Viability Assay

A bioluminescent method to kinetically monitor viability in cell culture up to 72 hours.

G9711, G9712, G9713

pHAb Reactive Dyes

Measure antibody internalization in real time with pH-sensitive fluorescent dyes.

G9841, G9831, G9835, G9845

Let's find the product that meets your needs.

Talk to a Scientist

Petra

Petra

Germany
Choose your country

Americas

Brazil
/
Canada
/
United States
/

Pacific Asia

Australia
China
/
India
Japan
/
Korea, Republic of
/
Singapore

Europe

Austria
/
Belgium
Denmark
(Incl. Faroe Isl. & Greenland)
Estonia
Finland
(Incl. Åland)
France
/
Germany
/
Iceland
Ireland
Italy
/
Luxembourg
Netherlands
Norway
(Incl. Svalbard)
Poland
/
Spain
/
Sweden
Switzerland
United Kingdom

Manage Cookies

Necessary Cookies

We use these cookies to ensure our site functions securely and properly; they are necessary for our services to function and cannot be switched off in our systems. They are usually only set in response to actions made by you which amount to a request for services, such as logging in, using a shopping cart or filling in forms. You can set your browser to block or alert you about these cookies, but some parts of our services will not work without them. Like the other cookies we use, strictly necessary cookies may be either first-party cookies or third - party cookies.

Preference Cookies

We use these cookies to remember your settings and preferences. For example, we may use these cookies to remember your language preferences.

Performance/Statistics Cookies

We use these cookies to collect information about how you interact with our services and to help us measure and improve them. For example, we may use these cookies to determine if you have interacted with a certain page.

Marketing Cookies

We and our advertising partners use these cookies to deliver advertisements, to make them more relevant and meaningful to you, and to track the efficiency of our advertising campaigns, both on our services and on other websites and social media.

Your Privacy Choices

Our website supports your privacy choices by allowing users in select regions to opt out of specific types of sharing/sales/use of cookies and technologies for Targeted Advertising. Visit our Privacy Policy and Cookie Policy pages to learn more about these topics.

Targeted Advertising

Deselect this checkbox to disable cookies related to targeted advertising in your Promega experience. These advertising cookies will no longer be set for this browser and device.

Note: These settings are based on necessary cookies that are placed on your device, and if those cookies are cleared or you access the website from another browser or device, advertising cookies might be reset by default in the future, from which you can opt-out.