LAG-3/MHCII Blockade Bioassay
Measure Potency and Stability of Biologics Designed to Block LAG-3 Interactions
- 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: JA1111
Catalog Number: JA1115
Catalog Number: JA1112
Catalog Number: K1150
Catalog Number: K1201
免疫チェックポイントシグナルを遮断するバイオロジックスの能力測定
LAG-3/MHCII Blockade Bioassay は、LAG-3とその最も特性化されたリガンドである主要組織適合性複合体 II (MHCII) との相互作用を遮断するためにデザインされた抗体あるいはその他のバイオロジックスの効力および安定性を測定する発光法による細胞ベースアッセイです。LAG-3 はCD223としても知られており、活性化 CD4+/ CD8+ T 細胞 および ナチュラルキラー (NK) 細胞に発現する免疫チェックポイント受容体です。 LAG-3 は腫瘍抗原および自己抗体に対する免疫反応を制御する重要な役割を果たしています。MHCII によるLAG-3への結合は TCRシグナル伝達、サイトカイン産生および活性化 T細胞の増殖を阻害します。LAG-3/MHCII 相互作用を遮断するためにデザインされた治療用抗体は、様々ながんに対する臨床試験で有望な結果を示しています。
LAG-3/MHCII Blockade Bioassay は LAG-3とそのリガンドである MHCII の相互作用を遮断するためにデザインされた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
The assay consists of two cell lines:
LAG-3 Effector Cells: Jurkat T cells stably expressing human LAG-3 and a luciferase reporter driven by T-cell activation pathway-dependent response elements.
MHC APC Cells: APC Cells presenting TCR Activating Antigen, a proprietary peptide designed to specifically activate the TCR on the LAG-3 Effector Cells.
When the two cell types are co-cultured in the presence of antigen, MHCII on the APC cells presents the Ag to the TCR on the LAG-3 Effector Cells to activate the Effector Cells. LAG-3 on the Effector Cells inhibits TCR signaling and promoter-mediated luciferase activity. Addition of the anti-LAG-3 antibody releases LAG-3 mediated inhibition, resulting in promoter-mediated luminescence.
The LAG-3/MHCII Blockade Bioassay is also available in CPM format, containing cryopreserved cells that can be thawed, propagated and banked for long-term use.
ワークフロー
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 LAG-3/MHCII Blockade Bioassay is a bioluminescent cell-based assay that overcomes the limitations of existing assays and can be used to measure the potency and stability of antibodies and other biologics targeting LAG-3.
Representation of the LAG-3/MHCII Blockade Bioassay. The bioassay consists of two cell lines, LAG-3 Effector Cells and MHCII APC Cells. When co-cultured, the LAG-3 inhibits TCR pathway-activated luminescence. The addition of anti-LAG-3 antibody blocks LAG-3 binding to MHCII, resulting in full TCR pathway activation, which can be detected in a dose-dependent manner by addition of Bio-Glo™ Reagent and quantitation with a luminometer.
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。LAG-3/MHCII Blockade 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 CTLA-4 Blockade Bioassay Shows Precision, Accuracy and Linearity.
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 94.4 | |
| 75 | 94.8 | |
| 100 | 100.3 | |
| 100 | 100.3 | |
| 200 | 99.8 | |
| Repeatability (% CV) | 100% (Reference) | 7.5 |
| Intermediate Precision (% CV) | 9.7 | |
| Linearity (r2) | 0.998 | |
| Linearity (y = mx + b) | y = 1.03x – 3.65 | |
| A 50–200% theoretical potency series of Control Ab, Anti-LAG-3, 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 LAG-3/MHCII Blockade Bioassay reflects the MOA and specificity of biologics designed to block the LAG-3/MHCII Interaction.
LAG-3 Effector Cells were incubated with TCR Activating Antigen and MHCII APC cells in the presence of anti-LAG-3, anti-PD-1, anti-CTLA-4 or anti-TIGIT antibodies, as indicated.
The LAG-3/MHCII Blockade Bioassay measures the activity of Control Ab, Anti-LAG-3.
MHCII APC Cells and TCR Activating Antigen were plated overnight. The following day, a titration of Control Ab, Anti-LAG-3 was added to the plate, followed by LAG-3 Effector Cells, and then incubated for 5 hours.
キットのフォーマットと関連製品
- LAG-3/MHCII Blockade Bioassay is available in 1X and 5X sizes.
- LAG-3/MHCII Blockade Bioassay is also available in CPM format (Cat.# JA1112).
- Control Ab, Anti-LAG-3, is available separately (Cat.# K1150).
- TCR Activating Antigen is available separately (Cat.# K1201).
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
LAG-3 Effector Cells |
J249A | 1 × 0.5ml | |
MHCII APC Cells |
J250A | 1 × 0.5ml | |
TCR Activating Antigen Stock Solution |
K120A | 1 × 60μl | View Product |
DMEM with 4.5g/L Glucose, L-Glutamine, Sodium Pyruvate |
J236A | 1 × 60ml | |
Fetal Bovine Serum |
J121A | 1 × 4ml | |
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
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 | Available Separately |
|---|---|---|---|
LAG-3 Effector Cells |
J249A | 5 × 0.5ml | |
MHCII APC Cells |
J250A | 5 × 0.5ml | |
TCR Activating Antigen Stock Solution |
K120A | 5 × 60μl | View Product |
DMEM with 4.5g/L Glucose, L-Glutamine, Sodium Pyruvate |
J236A | 5 × 60ml | |
Fetal Bovine Serum |
J121A | 5 × 4ml | |
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
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 | Available Separately |
|---|---|---|---|
LAG-3 Effector Cells (CPM) |
J245A | 2 × 1ml | |
TCR Activating Antigen Stock Solution |
K120B | 1 × 500μl | View Product |
MHCII APC Cells (CPM) |
J246A | 2 × 1ml |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
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-LAG-3 |
K115A | 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
See the Protocol for detailed storage conditions.
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
TCR Activating Antigen Stock Solution |
K120B | 1 × 500μl | View Product |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
See the Protocol for detailed storage conditions.
Resources
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