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PD-1/PD-L1 Blockade Bioassay

Part Numbers: J1250, J1255, J1252, J1191, J1195, J1201

J1250_PD-1-3aPD-L1-Blockade-Bioassay_3
J1252
J1250_PD-1-3aPD-L1-Blockade-Bioassay_3
customize-this-small

免疫チェックポイントバイオアッセイ

  • ICH ガイドラインに認証済み
  • 96- および 384-ウェルプレートフォーマット対応
  • 初代培養細胞は不要
  • 便利な ”融解 & アッセイ”だけの使い切りフォーマットと増殖が許容された cell propagation model (CPM) の2つのタイプ

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選択製品のカタログ番号: J1250

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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
Your price:
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.

アッセイの構成

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

標準化された試薬による最適な結果

  • ”解凍 ＆ アッセイ” だけのフォーマットは従来法に比べ飛躍的に時間と労力を削減し、バラつきも低減
  • キットは標準化されたフォーマットとして必要なすべての試薬を供給
  • ハイスループットワークフローにも容易に適応

アッセイの原理

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).

プロトコール

Complete Protocol

製品仕様

You are viewing: J1250 Change Configuration

選択製品の構成品内容

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

分析証明書

Search by lot number

使用制限

Not For Medical Diagnostic Use.

保存条件

Please see the Protocol for detailed storage conditions.

パテントおよび免責条項

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. 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.

製品仕様

You are viewing: J1255 Change Configuration

選択製品の構成品内容

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

分析証明書

Search by lot number

使用制限

Not For Medical Diagnostic Use.

保存条件

Please see the Protocol for detailed storage conditions.

パテントおよび免責条項

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. 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.

製品仕様

You are viewing: J1252 Change Configuration

選択製品の構成品内容

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

分析証明書

Search by lot number

使用制限

Not For Medical Diagnostic Use.

保存条件

Please see the Protocol for detailed storage conditions.

パテントおよび免責条項

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.

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.

製品仕様

You are viewing: J1191 Change Configuration

選択製品の構成品内容

Item Part # Size

aAPC/CHO-K1 Cells

 J119A 1 × 0.5ml

SDS

Search for SDS

分析証明書

Search by lot number

使用制限

Not For Medical Diagnostic Use.

保存条件

XX

Please see the Protocol for detailed storage conditions.

パテントおよび免責条項

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.

製品仕様

You are viewing: J1195 Change Configuration

選択製品の構成品内容

Item Part # Size

aAPC/CHO-K1 Cells

 J119A 5 × 0.5ml

SDS

Search for SDS

分析証明書

Search by lot number

使用制限

Not For Medical Diagnostic Use.

保存条件

XX

Please see the Protocol for detailed storage conditions.

パテントおよび免責条項

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.

製品仕様

You are viewing: J1201 Change Configuration

選択製品の構成品内容

Item Part # Size

Control Ab, Anti-PD-1

 J120A 1 × 100μg

SDS

Search for SDS

分析証明書

Search by lot number

使用制限

Not For Medical Diagnostic Use.

保存条件

CC

Please see the Protocol for detailed storage conditions.

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