FcγRIIa-H ADCP Bioassay
Measure Potency and Stability of Biologics that Bind and Activate ADCP through FcγRIIa-H
- Complete format contains everything you need to get started
- Use Core format with customer-defined Ab and target cells
- Also available in cell propagation model (CPM) format
Catalog Number:
Choose a Product
Size
Catalog Number: G9901
Catalog Number: G9991
Catalog Number: G9995
Catalog Number: G9871
Measure Potency and Stability of Biologics that Bind and Activate FcyRIIa-H
抗体依存性細胞貪食 (ADCP) は治療用抗体の重要な作用機序 (MOA: mechanism of action) であり、In vivoではADCPはFcγRIIa、FcγRIおよびFcγRIIIa を介して単球、マクロファージ、好中球、樹状細胞により媒介されます。3つすべての受容体がADCPに関与しますが、FcγRIIaはこのプロセスに最も重要なFcγ受容体であると考えられています。
FcγRIIa-H ADCP Reporter Bioassay は生物学的に関連性のあるMOAベースのアッセイで、FcγRIIaに特異的に結合し、活性化する抗体やその他のバイオロジックスの力価や安定性測定に使用することができます。本アッセイに含まれるJurkat細胞はヒト FcγRIIa-H (高親和性 H131 バリアント) を安定に発現し、NFAT-誘導性ルシフェラーゼが含まれます。
このバイオアッセイは、ICHガイドラインに準じた事前承認がなされており、力価および安定性の試験におけるルーチンな使用に要求される精度 (precision) と 正確度 (accuracy) および直線性を示します。また、このバイオアッセイのワークフローは従来のプライマリー細胞ベースのアッセイとは異なり、シンプルかつ頑健で96/384 ウェルプレートフォーマットに適合し、品質管理された薬剤開発セッティングにそのまま利用することができます。
キットの構成
FcγRIIa-H ADCP Reporter Bioassayは複数の製品キットからお選びいただけます:
Complete Kit （スターターキット）
- 構成品： FcγRIIa-H Effector Cells, Target Cells (Raji), Control Antibody, Cell Culture Medium および Assay Reagent
Core Kits （自前の抗体および標的細胞を用いたルーチンでの使用に推奨）
- 構成品： FcγRIIa-H Effector Cells, Cell Culture Medium および Assay Reagent
- 1X と 5X の2サイズを用意
- FcγRIIa を介したADCP作用機序の証明と確認
- 抗体の力価および安定性の測定
- 抗体のグリコシル化およびアフコシル化の機能への影響を評価
- 抗体のスクリーニング
FcγRIIIa-H ADCP Bioassay Effector Cells, Propagation Modelは FcγRIIa-H Effector Cellsの増殖、バンキングを行うことができます。 Bio-Glo™ Luciferase Assay Systemは FcγRIIa-H ADCP Bioassay Effector Cells, Propagation Modelを使用する場合に必要な試薬です。
【ライセンス】ADCP Bioassay
- Richards, J.O. et al. (2008) Optimization of antibody binding to FcγRIIa enhances macrophage phagocytosis of tumor cells. Mol. Cancer Ther. 7, 2517–27.
- Dugast, A.S. et al. (2011) Decreased Fc-receptor expression on innate immune cells is associated with impaired antibody mediated cellular phagocytic activity in chronically HIV-1 infected individuals. Virology 415, 160–7.
- Ackerman, M.E. et al. (2013) Enhanced phagocytic activity of HIV-specific antibodies correlates with natural production of immunoglobulins with skewed affinity for FcγR2a and FcγR2b. J. Virol. 87, 5468–76.
- Tebo, A.E. et al. (2002) Fcγ receptor-mediated phagocytosis of Plasmodium falciparum-infected erythrocytes in vitro. Clin. Exp. Immunology 130, 300–6.
ワークフロー
標準化された試薬による改善点
- 製品に含まれる細胞を融解して使用するだけなので、従来法よりも飛躍的に時間と労力が低減し、バラつきも減少
- キットは標準化フォーマットとして必要な全ての試薬を供給
- ハイスループットなワークフローにも容易に適応
The Assay Principle
The Fc Effector Reporter Bioassays use an alternative readout from traditional primary cell based assays: the FcγR and NFAT-mediated activation of luciferase activity in the effector cells. Primary cells are replaced with a Jurkat cell line stably expressing human FcγRIIa H-131and NFAT-induced luciferase.
Luminescent Assay Monitors Fc Effector Activity
Following engagement with the Fc region of a relevant antibody bound to a Target Cell, ADCP Bioassay Effector Cells expressing FcγRIIa-H131 with transduce intracellular signals resulting in NFAT-RE-mediated luciferase activity that can be easily quantified.
Assay Performance Data
Accuracy and Precision are critical to bioassays and the development of new biologics. The ADCP Reporter Bioassays are both accurate and precise and have been prequalified according to ICH guidelines and show the precision, accuracy and linearity required for routine use in potency and stability studies. Accuracy is a measure of how close results are to a target or reference value. Precision is a measure of repeatability.
Accuracy: Measure of ability to replicate 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.
ADCP Reporter Bioassay Shows Precision, Accuracy and Linearity
Raji Target Cells
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 99.1 | |
| 75 | 102.7 | |
| 150 | 105.0 | |
| 200 | 99.1 | |
| Repeatability (% CV) | 100% (Reference) | 6.5 |
| Intermediate Precision (% CV) | 3.5 | |
| Linearity (r2) | 0.997 (50–200% range) | |
| Linearity (y = mx + b) | y = 0.997x + 1.95 | |
|
A 50–200% theoretical relative potency series of the ADCP Control Ab, anti-CD20 (IgG1), was analyzed in triplicate in three independent experiments performed on three days. Luciferase activity was quantified using the Bio-Glo™ Reagent. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells. |
||
The ADCP Bioassay reflects the MOA and specificity of antibodies designed to bind and activate FcγRIIa-H
Product Format
Complete Kit
- Includes FcγRIIa-H Effector Cells, Target Cells (Raji), Control Antibody, Cell Culture Medium and Assay Reagents.
- Recommended for use as a starter kit.
Core Kit
- Includes FcγRIIa-H Effector Cells, Cell Culture Medium and Assay Reagents.
- Recommended for routine use with customer-defined antibody and target cells.
- Available in 1X and 5X sizes.
Cell Propagation Model
- Includes FcγRIIa-H Effector Cells (CPM); these are cryopreserved cells that can be thawed, propagated and banked for long-term use.
- Use to measure the potency and stability of antibodies and other biologics with Fc domains that specifically bind and activate FcγRIIa.
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
FcγRIIa-H Effector Cells |
G988A | 1 × 0.62ml |
Target Cells (Raji) |
G960A | 1 × 0.55ml |
ADCP Control Ab, Anti-CD20 |
G989A | 1 × 2.5μg |
Low IgG Serum |
G711A | 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 |
|---|---|---|
FcγRIIa-H Effector Cells |
G988A | 1 × 0.62ml |
Low IgG Serum |
G711A | 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 |
|---|---|---|
FcγRIIa-H Effector Cells |
G988A | 5 × 0.62ml |
Low IgG Serum |
G711A | 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 |
|---|---|---|
FcγRIIa-H Effector Cells (CPM) |
G987A | 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.
Resources
No related resources available
Related Products
類似製品
ADCC Reporter Bioassay, F Variant
Fc エフェクター機能における FcγRIIa F158 多型の影響を定量するための作用機序ベースのレポーターバイオアッセイ
G9790, G9798, G9302
ADCC Reporter Bioassay, V Variant
高親和性ヒトFcγRIIIa-V受容体を介したADCC活性を持つ抗体の力価および安定性を評価
G7015, G7014, G7010, G7016, G7013, G7018, G7102, GA1130
PD-1/PD-L1 Blockade Bioassay
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力価測定に
J1250, J1255, J1252, J1191, J1195, J1201
Lumit® FcγR Binding Immunoassays
ヒトFc受容体と抗体またはFc融合タンパク質との相互作用を測定するための新規ホモジニアス競合アッセイ
W7030, W7031, W7040, W7041, W7050, W7051, W7060, W7061, W7070, W7071, W7080, W7081
