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
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Catalog Number: G7015
Catalog Number: G7014
Catalog Number: G7010
Catalog Number: G7016
Catalog Number: G7013
Catalog Number: G7018
Catalog Number: G7102
Catalog Number: GA1130
Eliminate the Uncertainty and Variability of Primary Cell Based ADCC Assays
Fc receptor-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) is an important mechanism of action (MOA) by which antibodies target diseased cells for elimination. The human FcyRIIIa gene displays a polymorphism in the position coding for amino acid residue 158. This translates to a higher affinity FcyRIIIa variant having a valine at amino acid 158 (V158) and a lower affinity FcyRIIIa variant carrying phenylalanine F158.
Classic methods used to measure ADCC use primary donor peripheral blood mononuclear cells (PBMCs) or purified natural killer (NK) cells that express Fc receptors on their cell surface. Killing of target cells is an endpoint of this pathway activation and is used in classic ADCC bioassays, which rely on donor peripheral blood mononuclear cells (PBMCs) or the natural killer (NK) cell subpopulation as effector cells isolated from blood donors.
These primary cell-based assays are highly variable in response, are tedious to prepare and can result in high background readings. The ADCC Reporter Bioassays overcome many of the limitations of traditional methods by eliminating the requirement for primary cell culture.
The Workflow
Standardized Reagents Improve Results
- Thaw-and-use cell 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.
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 FcγRIIIa V158 and NFAT-induced luciferase.
Luminescent Assay Monitors Fc Effector Activity
Following engagement with the Fc region of a relevant antibody bound to a Target Cell, ADCC Bioassay Effector Cells expressing FcγRIIIa will transduce intracellular signals resulting in NFAT-mediated luciferase activity that can be easily quantified.
Assay Performance Data
Accuracy and Precision are critical to bioassays and development of biologics. The ADCC Reporter Bioassays are both accurate and precise. 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.
Product Format
Complete Kits
- Include ADCC Bioassay Effector Cells, Target Cells, Control Antibody, Cell Culture Medium and Assay Reagents.
- Recommended for use as a starter kit.
- Available with either Raji or WIL2-S Target Cells.
Core Kits
- Include ADCC Bioassay 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.
Target Kits
- Include ADCC Bioassay Target Cells and Control Antibody.
- Recommended for use as a control with all Fc Effector Bioassay Core Kits.
- Available with either Raji or WIL2-S Target Cells.
ADCC Bioassay Effector Cells, Propagation Model
- Includes ADCC Bioassay Effector Cells (CPM); these are cryopreserved cells that can be thawed, propagated and banked for long-term use. Product includes two vials at 2 × 107 cells/ml and 1.0ml/vial. One vial should be thawed, propagated and cells frozen to create a cell bank. The remaining vial should be reserved as backup.
- Bio-Glo™ Luciferase Assay System is the required reagent for use with ADCC Bioassay Effector Cells, Propagation Model.
Control Antibody, Anti-CD20
- Intended for use as a control antibody with the ADCC Bioassay.
For additional format options please enquire.
ADCC Reporter Bioassay Precision, Accuracy and Linearity
WIL2-S Target Cells
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 97.7 | |
| 75 | 88.5 | |
| 125 | 98.4 | |
| 150 | 98.4 | |
| Repeatability (% CV) | 100% (Reference) | 5.0% |
| Intermediate Precision (% CV) | 7.3% | |
| Linearity (r2) | 0.995 (50–200% range) | |
| Linearity (y = mx + b) | y = 1.016x – 0.052 | |
Raji Target Cells
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 101.0 | |
| 75 | 101.2 | |
| 150 | 93.8 | |
| 200 | 96.7 | |
| Repeatability (% CV) | 100% (Reference) | 9.1 |
| Intermediate Precision (% CV) | 3.0 | |
| Linearity (r2) | 0.997 (50–200% range) | |
| Linearity (y = mx + b) | y = 0.922x + 5 | |
ADCC activity is a major mode of action of therapeutic monoclonal antibodies and is generally determined using a cell-based assay with primary PBMCs. ADCC Reporter Bioassay allows to measure the potency of an antibody without the constraint of PBMC donors’ variability and with a high level of performance. This assay was successfully validated and implemented in our company for batch release testing.
Dr. Fabian Vandermeers, Sr R&D Technical Leader, Quality Assistance S.A.
Protocols
Complete Protocol
Specifications
Catalog Number:
What's in the box?
| 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 |
SDS
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Storage Conditions
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 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.
What's in the box?
| 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 |
SDS
Search for SDSCertificate of Analysis
Storage Conditions
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 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.
What's in the box?
| 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 |
SDS
Search for SDSCertificate of Analysis
Storage Conditions
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 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.
What's in the box?
| Item | Part # | Size | Concentration |
|---|---|---|---|
ADCC Bioassay Target Cells (Raji) |
G870A | 1 × 0.55ml | |
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
SDS
Search for SDSCertificate of Analysis
Storage Conditions
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 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.
What's in the box?
| Item | Part # | Size | Concentration |
|---|---|---|---|
ADCC Bioassay Target Cells (WIL2-S) |
G727A | 1 × 0.55ml | |
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
SDS
Search for SDSCertificate of Analysis
Storage Conditions
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 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.
What's in the box?
| 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 |
SDS
Search for SDSCertificate of Analysis
Storage Conditions
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 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.
What's in the box?
| Item | Part # | Size |
|---|---|---|
ADCC Bioassay Effector Cells (CPM) |
G701B | 2 × 1.0ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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.
What's in the box?
| Item | Part # | Size | Concentration |
|---|---|---|---|
Control Ab, Anti-CD20 |
GA113A | 1 × 5μg | 0.5mg/ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
Please see the Protocol for detailed storage instructions.
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