ADCC Reporter Bioassay, V Variant
MOA-Based Bioassay to Measure FcγRIIIa (V158) Function
- Complete Kit: Everything you need to get started
- Core Kit: Used with customer-defined Ab and target cells
- Target Kit: Control used with Core Kit
- Also available in cell propagation model (CPM) format
Catalog Number:
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Size
Catalog Number: G7015
Catalog Number: G7014
Catalog Number: G7010
Catalog Number: G7016
Catalog Number: G7013
Catalog Number: G7018
Catalog Number: G7102
Catalog Number: GA1130
プライマリー細胞ベースのADCCアッセイの不確実性とバラつきを解消
Fc受容体を介した抗体依存性細胞傷害（ADCC：Antibody-dependent cell-mediated cytotoxicity）は、抗体が罹患細胞を標的として排除するための重要な作用機序（MOA）です。ヒトFcγRIIIa遺伝子は158番目のアミノ酸残基をコードする位置に多型を示します。これにより、158番目のアミノ酸がバリン（V）である高親和性FcγRIIIaバリアント（V158）と、フェニルアラニン（F）を持つ低親和性バリアント（F158）が生じます。
従来のADCC測定には、細胞表面にFc受容体を発現するドナー由来の末梢血単核球（PBMC）または精製ナチュラルキラー（NK）細胞が使用されてきました。標的細胞の傷害がこの経路活性化のエンドポイントとなりますが、これらのプライマリー細胞ベースのアッセイはアッセイ間のバラつきが大きく、調製が煩雑で、バックグラウンドが高くなることが課題でした。
ADCC Reporter Bioassayはプライマリー細胞培養を不要にすることで、従来法の多くの課題を克服しています。
【ライセンス】ADCC Bioassay 【Ｑ＆Ａ】ADCC Reporter Bioassay
【かわら版】驚きの再現性 : 発光レポーターを利用した ADCC アッセイ
【かわら版】ADCC 発光レポーターバイオアッセイを用いた 抗 TNF 抗体医薬品および抗ウイルスワクチンの評価
ワークフロー
標準化された試薬による結果の改善
- Thaw-and-Use形式の細胞により、従来法と比べて大幅な時間と労力の削減を実現し、バラつきも低減
- 標準化フォーマットで必要な試薬をすべてキットに同梱
- ハイスループットワークフローにも容易に対応
アッセイ原理
このFc Effector Reporter Bioassayは、従来のプライマリー細胞ベースのアッセイとは異なる検出法を採用しています。エフェクター細胞内でFcγRおよびNFATを介したルシフェラーゼ活性の活性化を検出します。プライマリー細胞は、ヒトFcγRIIIa V158およびNFAT誘導性ルシフェラーゼを安定に発現するJurkat細胞株に置き換えられています。
発光アッセイによる Fc エフェクター活性のモニタリング
標的細胞に結合した抗体のFc領域がADCC Bioassay Effector Cellsに認識されると、FcγRIIIaを介した細胞内シグナルが伝達され、NFAT誘導性ルシフェラーゼ活性として容易に定量できます。
アッセイパフォーマンスデータ
正確度（Accuracy）と精度（Precision）は、バイオアッセイおよびバイオロジクス開発において非常に重要です。ADCC Reporter Bioassayはこの両方を兼ね備えています。
- 正確度：参照値または「標的」値をどれだけ忠実に再現できるかの指標
- 再現性：同一条件下（同一分析者・同一ラボ・同日）で同一均質サンプルを複数回アッセイした際の再現性の指標
- 中間精度：異なる分析者・機器・日時で同一均質サンプルを複数回アッセイした際の再現性の指標
- 直線性：サンプル濃度に比例した測定値が得られるかどうかの指標
製品フォーマット
Complete Kits
- ADCC Bioassayエフェクター細胞、標的細胞、コントロール抗体、細胞培養培地およびアッセイ試薬で構成
- スターターキットとしての使用を推奨
- RajiまたはWIL2-S標的細胞を用意
Core Kits
- ADCC Bioassayエフェクター細胞、細胞培養培地およびアッセイ試薬で構成
- お客様がご用意する抗体および標的細胞を用いたルーチン実験での使用を推奨
- 1Xと5Xサイズを用意
Target Kits
- ADCC Bioassay標的細胞およびコントロール抗体で構成
- すべてのFc Effector Bioassay Core Kitsのコントロールとしての使用を推奨
- RajiまたはWIL2-S標的細胞を用意
ADCC Bioassay Effector Cells, Propagation Model
- ADCC Bioassayエフェクター細胞（CPM）を含む凍結保存細胞です。解凍・増殖後にバンキングして長期使用が可能です。2バイアル入り（2×10⁷個/ml、1.0ml/バイアル）。1バイアルを解凍・増殖させ凍結して細胞バンクを作製し、残りの1バイアルはバックアップとして保管してください。
- 本製品の使用にはBio-Glo™ Luciferase Assay Systemが必要です。
Control Antibody, Anti-CD20
- ADCC Bioassayのコントロール抗体として使用します。
その他のフォーマットオプションについてはお問い合わせください。
ADCC Reporter Bioassay 正確度・精度・直線性
WIL2-S 標的細胞
| パラメーター | 結果 | |
|---|---|---|
| 正確度 | 期待相対力価（%） | 回収率（%） |
| 50 | 97.7 | |
| 75 | 88.5 | |
| 125 | 98.4 | |
| 150 | 98.4 | |
| 再現性 (% CV) | 100% (Reference) | 5.0% |
| 中間精度 (% CV) | 7.3% | |
| 直線性 (r2) | 0.995 (50–200% 範囲) | |
| 直線性 (y = mx + b) | y = 1.016x – 0.052 | |
Raji 標的細胞
| パラメーター | 結果 | |
|---|---|---|
| 正確度 | 期待相対力価（%） | 回収率（%） |
| 50 | 101.0 | |
| 75 | 101.2 | |
| 150 | 93.8 | |
| 200 | 96.7 | |
| 再現性 (% CV) | 100%（基準値） | 9.1 |
| 中間精度 (% CV) | 3.0 | |
| 直線性 (r2) | 0.997 (50–200% 範囲) | |
| 直線性 (y = mx + b) | y = 0.922x + 5 | |
ADCCはモノクローナル抗体医薬品の主要な作用機序のひとつであり、一般的にはプライマリーPBMCを用いた細胞ベースのアッセイで評価されます。ADCC Reporter Bioassayにより、PBMCドナーのバラつきに制約されることなく、高い性能で抗体の力価を測定できます。このアッセイは当社で適切にバリデーションされ、ロット出荷試験に導入されています。
Quality Assistance S.A. シニアR&D技術リーダー Fabian Vandermeers博士
Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
ADCC Bioassay Effector Cells |
G701A | 1 × 1 vial | |
ADCC Bioassay Target Cells (Raji) |
G870A | 1 × 0.55ml | |
Low IgG Serum |
G711A | 1 × 4ml | |
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
RPMI 1640 Medium |
G708A | 1 × 36ml | |
Bio-Glo™ Luciferase Assay Buffer |
G719A | 1 × 10ml | |
Bio-Glo™ Luciferase Assay Substrate |
G720A | 1 × 1 vial |
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Effector Cells or the luciferase gene stably transfected within the Effector Cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all determinations of luminescence activity of 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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Target 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 | Concentration |
|---|---|---|---|
ADCC Bioassay Effector Cells |
G701A | 1 × 1 vial | |
ADCC Bioassay Target Cells (WIL2-S) |
G727A | 1 × 0.55ml | |
Low IgG Serum |
G711A | 1 × 4ml | |
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
RPMI 1640 Medium |
G708A | 1 × 36ml | |
Bio-Glo™ Luciferase Assay Buffer |
G719A | 1 × 10ml | |
Bio-Glo™ Luciferase Assay Substrate |
G720A | 1 × 1 vial |
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Effector Cells or the luciferase gene stably transfected within the Effector Cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all determinations of luminescence activity of 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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Target 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 |
|---|---|---|
ADCC Bioassay Effector Cells |
G701A | 1 × 1 vial |
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 |
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Effector Cells or the luciferase gene stably transfected within the Effector Cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all determinations of luminescence activity of 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 |
|---|---|---|---|
ADCC Bioassay Target Cells (Raji) |
G870A | 1 × 0.55ml | |
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Target 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.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
ADCC Bioassay Target Cells (WIL2-S) |
G727A | 1 × 0.55ml | |
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Target 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.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
ADCC Bioassay Effector Cells |
G701A | 5 × 1 vial |
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 |
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Effector Cells or the luciferase gene stably transfected within the Effector Cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all determinations of luminescence activity of 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 |
|---|---|---|
ADCC Bioassay Effector Cells (CPM) |
G701B | 2 × 1.0ml |
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 instructions.
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) drug discovery and development of biologic drugs, (3) quality assurance testing of biologic drugs, and (4) product release assays for biologic drugs. 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 Effector Cells or the luciferase gene stably transfected within the Effector Cells, except that recipient may propagate and store Effector Cells for use in Antibody-Dependent Cell-Mediated Cytotoxicity assays. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all determinations of luminescence activity of 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-CD20 |
GA113A | 1 × 5μg | 0.5mg/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 instructions.
Resources
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