GITR Bioassay
Measure Potency and Stability of Biologics Designed to Activate GITR
- Prequalified according to ICH guidelines
- 96- and 384-well plate formats
- Supplied in convenient thaw-and-use format; also available in cell propagation model (CPM) format
Catalog Number:
Choose a Product
Size
Catalog Number: JA2291
Catalog Number: JA2295
Catalog Number: J2272
Catalog Number: K1171
安定した結果が得られるシンプルなバイオアッセイ
GITRバイオアッセイは、GITRに結合して活性化させるリガンドまたはアゴニスト抗体の力価と安定性を測定するために使用される生物発光細胞ベースのアッセイです。腫瘍壊死因子（TNF）受容体スーパーファミリーのメンバーであるGITR（CD357 / TNFRSF18）は、ほとんどの免疫細胞で広く発現し、活性化T細胞をさらに上方制御する共刺激受容体です。細胞表面でGITRリガンド（GITRL）がGITRに作用すると、T細胞の増殖と、インターロイキン-2（IL-2）およびIL-9を含むサイトカインの産生を促進します。
GITRバイオアッセイは、GITRを活性化するように設計されたバイオロジックス（生物製剤）の作用機序（MOA）を反映しています。
Assay Components
The thaw-and-use format of the GITR Bioassay (Cat.# JA2291, JA2295) offers convenience and minimizes variability. The assay consists of a genetically engineered cell line, GITR Effector Cells, that expresses human GITR and a luciferase reporter driven by a response element that can respond to GITR ligand/agonist antibody stimulation.Based on the antibody properties to be tested, the GITR Bioassay can be conducted in either single-cell systems or with FcγRIIb CHO-K1 Cells (Cat.# JA2251, JA2255) to determine the agonist antibodies that can activate GITR in a FcγR-dependent manner.
The GITR Effector Cells are also available in Cell Propagation Model format (CPM; Cat.# J2272), as cryopreserved cells that can be thawed, propagated and banked for long-term use.
Workflow
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 GITR Bioassays measure the potency and stability of ligands or agonist antibodies and other biologics that bind and activate GITR.
Panel A. Assay with FcγRIIb-dependent agonist antibody. The bioassay consists of two engineered cell lines, GITR Effector Cells and FcγRIIb CHO-K1 Cells. In the presence of FcγRIIb CHO-K1 Cells, the anti-GITR antibody can be crosslinked, thereby inducing GITR pathway-activated luminescence.
Panel B. Assay with FcγRIIb-independent agonist antibody or ligand. The bioassay consists of one engineered cell line, GITR Effector Cells. In the absence of agonist antibody or GITR ligand, the GITR receptor is not activated and luminescence signal is low. The addition of agonist antibody or GITR ligand induces the GITR pathway-activated luminescence, which can be detected in a dose-dependent manner.
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.
The GITR Bioassays Show Precision, Accuracy and Linearity.
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 99.3 | |
| 75 | 98.9 | |
| 125 | 102.5 | |
| 150 | 102.9 | |
| Repeatability (% CV) | 100% (Reference) | 1.3 |
| Intermediate Precision (% CV) | 6.6 | |
| Linearity (r2) | 0.997 | |
| Linearity (y = mx + b) | y = 1.053x – 3.742 | |
| A 50–150% theoretical potency series of Control Ab, Anti-GITR 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. | ||
Product Kit Formats and Related Products
- GITR Bioassay is available in 1X and 5X sizes.
- Control Ab, Anti-GITR (Cat.# K1171) is available separately.
- FcγRllb CHO-K1 Cells (Cat.# JA2251, JA2255) are available separately.
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
GITR Effector Cells |
J229A | 1 × 0.5ml |
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
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 |
|---|---|---|
GITR Effector Cells |
J229A | 5 × 0.5ml |
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
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 |
|---|---|---|
GITR Effector Cells (CPM) |
J227A | 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-GITR |
K117A | 1 × 50μg | 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.
Resources
No related resources available
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