PD-1+TIGIT Combination Bioassay
Measure potency and stability of biologics designed to block immune checkpoint signals
- 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
Catalog Number:
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Catalog Number: J2211
Catalog Number: J2215
Catalog Number: J2102
Catalog Number: J2051
Catalog Number: J1921
免疫チェックポイントのシグナルを遮断するバイオロジックスの力価測定
PD-1およびTIGIT はどちらも T細胞上に発現する免疫チェックポイント受容体です。近接する細胞上のリガンドPD-L1がPD-1に結合するとTCR シグナリング、TCRを介した細胞増殖、転写活性化およびサイトカインの生産が阻害されます。TIGIT のリガンドはCD155で、TIGIT はCD155への結合に対してCD226と競合あるいはCD226 のホモ二量体化に直接干渉することにより、NK細胞死、T細胞活性化を負に調節します。
前臨床がん研究ではPD-1 と TIGIT の両方の遮断はどちらか一方のパスウェイ遮断よりも優れていることが示唆されています。本 PD-1+TIGIT コンビネーションバイオアッセイは PD-1/PD-L1 と TIGIT/CD155 の各相互作用を遮断するバイオロジックスの作用機序 (MOA)を反映しています。
アッセイの構成
本アッセイは遺伝子改変された2種類の細胞株で構成されています:
PD-1+TIGIT Effector Cells: ヒトPD-1、 TIGIT および TCR 活性化および CD226 共刺激の両方に応答するネイティブプロモーターより駆動するルシフェラーゼレポーター を発現する Jurkat T 細胞
PD-L1+CD155 aAPC/CHO-K1 Cells: ヒト PD-L1 、CD155 とともに抗原非依存的にTCR 複合体を活性化するために遺伝子改変された細胞表面タンパク質を発現する CHO-K1 細胞
2種類の細胞種を共培養した場合、PD-1はPD-L1に結合してTCR誘導性活性化およびプロモーターを介した発光を阻害します。抗 PD-1 Ab または抗 TIGIT Ab の添加はそれぞれPD-1と PD-L1 または TIGIT と CD155 の相互作用を遮断し、プロモーターを介した発光が強くなります。PD-1/PD-L1 および TIGIT/CD155 の両方の相互作用を遮断する抗体の場合は 2つのパスウェイの相乗効果により非常に強い発光を生じます。
ワークフロー
従来法 （プライマリー細胞を使用）
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
PD-1+TIGIT Combination Bioassay は従来のアッセイ法の限界を超えて、PD-1 および TIGIT を標的とする抗体やその他のバイオロジックスの力価や安定性の測定に使用できる発光法による細胞ベースのアッセイです。
Representation of the PD-1+TIGIT Combination Bioassay. The bioassay consists of two genetically engineered cell lines, PD-1+TIGIT Effector Cells and PD-L1+CD155 aAPC/CHO-K1 Cells. Panel A. When co-cultured, PD-1 inhibits TCR pathway-activated luminescence, and TIGIT inhibits CD226 pathway-activated luminescence. The addition of anti-PD-1 Ab blocks PD-1 binding to PD-L1, resulting in full TCR pathway activation. The addition of anti-TIGIT Ab blocks the TIGIT/CD155 interaction, thereby re-establishing CD226 pathway-activated luminescence. Blocking of PD-1/PD-L1 and TIGIT/CD155 can be detected in a dose-dependent manner by addition of Bio-Glo™ Reagent and quantitation with a luminometer. Panel B. When co-cultured with Effector Cells that do not express PD-1 or TIGIT (Cat.# J1920), TCR activation and CD226/CD155 induce luminescence.
アッセイパフォーマンスデータ
正確度 (accuracy) と 精度 (precision) はバイオアッセイやバイオロジックス開発において非常に重要です。PD-1 + TIGIT Combination Bioassay は正確度と 精度を兼ね備えたアッセイです（正確度とは標的または参照値に対してどれほど近かったかという度合いであり、精度は再現性の度合いです）。本アッセイは正確度 と 精度 のためのICHガイドラインに従い開発、事前承認されており、抗体医薬の研究開発に使用するバイオアッセイの様々なアプリケーションに必要とされるパフォーマンス特性を備えています。
The PD-1+TIGIT Combination Bioassay Shows Precision, Accuracy and Linearity.
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 95.3 | |
| 70 | 105.5 | |
| 140 | 104.9 | |
| 200 | 109.9 | |
| Repeatability (% CV) | 100% (Reference) | 4.1 |
| Intermediate Precision (% CV) | 10.4 | |
| Linearity (r2) | 0.999 | |
| Linearity (y = mx + b) | y = 1.131x + 0.028 | |
| A 50–200% theoretical potency series of a 1:1 ratio of nivolumab (anti-PD-1 Ab) and Control Ab, Anti-TIGIT, was analyzed in triplicate in three independent experiments performed on three days by two analysts. Bio-Glo™ Reagent was added and luminescence quantified using the GloMax® Discover Detection System. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells. | ||
The PD-1+TIGIT Combination Bioassay reflects the mechanism of action (MOA) and specificity of biologics designed to block the PD-1/PD-L1 and TIGIT/CD155 interaction.
The PD-1+TIGIT Combination Bioassay measures the activity of Control Ab, Anti-PD-1, and Control Ab, Anti-TIGIT.
キットのフォーマットと関連製品
- 1X および 5X サイズより選択可能
- Control Ab, Anti-PD-1, Anti-TIGIT の別売品
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
PD-1+TIGIT Effector Cells |
J176A | 1 × 0.5ml |
PD-L1+CD155 aAPC/CHO-K1 Cells |
J184A | 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
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
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.
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.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
PD-1+TIGIT Effector Cells |
J176A | 5 × 0.5ml |
PD-L1+CD155 aAPC/CHO-K1 Cells |
J184A | 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
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
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.
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.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
PD-1+TIGIT Effector Cells (CPM) |
J172A | 2 × 1ml |
PD-L1+CD155 aAPC/CHO-K1 Cells (CPM) |
J183A | 2 × 1ml |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
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.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Control Ab, Anti-TIGIT |
J205A | 1 × 100μg | 1mg/ml |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
The transfer of this product is conditioned on the buyer using the purchased product for (i) research use, (ii) discovery, development and monitoring of biologic drugs and vaccines, (iii) quality assurance and quality control testing of biologic drugs and vaccines, and (iv) product release assays for biologic drugs and vaccines conducted by the buyer, including contract research or any fee for service research, and the buyer must not (1) use this product or its components for (1a) any use, indirectly (except as allowed in (iii) and (iv) above) or directly in manufacturing, or (1b) diagnostic, therapeutic or prophylactic purposes; and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
TCR/CD3 Effector Cells (IL-2) |
J129A | 1 × 0.8ml |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
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.
