ADCP Reporter Bioassay (THP-1)
Measure Potency and Stability of Biologics that Bind and Activate ADCP through FcγRs
- 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:
Choose a Product Format
Size
Catalog Number: JA9411
Catalog Number: JA9415
Catalog Number: GA1272
Catalog Number: GA6020
Simple Bioassay, Quality Results
Antibody-dependent cellular phagocytosis (ADCP) is an important mechanism-of-action (MOA) for antibody-based immunotherapies. ADCP can be mediated by monocytes, macrophages, neutrophils and dendritic cells via FcγRIIa, FcγRI and FcγRIIIa. In myeloid cells, the expression level of the various receptors is highly dynamic and influenced by cell lineage, tissue microenvironment and local inflammatory state. These receptors can participate in antibody recognition, receptor clustering and signaling events that result in ADCP.
Current methods for measuring ADCP rely on the isolation of primary human monocytes, ex vivo differentiation into macrophages and direct measurement of phagocytosis. These assays are laborious and highly variable due to their reliance on donor cells, complex assay protocols and unqualified assay reagents. As a result, these assays are difficult to establish in quality-controlled, drug development settings.
The ADCP Reporter Bioassay (THP-1) is a bioluminescent reporter cell-based assay used to measure the potency and stability of antibodies and other biologics with Fc domains that bind and activate FcγRs.
Bioassay Components and Product Formats
The ADCP Reporter Bioassay consists of a genetically engineered cell line:
ADCP Effector Cells (THP-1): A THP-1 monocyte cell line with endogenous expression of FcγRs. This cell line has been genetically engineered to express a NanoLuc® (NL) luciferase reporter driven by FcγR-dependent response elements.
Thaw-and-Use Kits (1X, 5X):
- Include ADCP Effector Cells (THP-1), cell culture media and assay reagents
- Recommended for routine use with customer-defined antibody and target cells
- Available in 1X and 5X sizes
Cell Propagation Model format (CPM; Cat.# GA1272):
- Includes the ADCP Effector Cells (THP-1) (CPM); these are cryopreserved cells that can be thawed, propagated and banked for long-term use.
- Use with Bio-Glo-NL™ Luciferase Assay System
Cell Bank format (Cat.# GA6020):
- Available for customers who are planning to expand and bank their CPM purchase, but they would like to use the cell expansion service offered by Promega.
When co-cultured with a target cell and relevant antibody, the ADCP Reporter Bioassay Effector Cells (THP-1) bind the Fc domain of the antibody, resulting in promoter-driven luminescence.
Workflow
Standardized Reagents Improve Results
- Thaw-and-use format provides significant time and labor savings over traditional methods while reducing variability.
- Easily amenable to high-throughput workflows.
Assay Principle
The ADCP Reporter Bioassay (THP-1) reflects the MOA of biologics designed to bind and activate FcγRs.
Representation of the ADCP Reporter Bioassay (THP-1). The bioassay consists of a genetically engineered cell line, ADCP Reporter Bioassay Effector Cells (THP-1), an antigen-expressing target cell and an antigen-specific antibody. When all components are co-cultured, the antibody simultaneously binds target cell antigen and FcγRs on the surface of the effector cells. This results in receptor clustering, intracellular signaling and luciferase activity.
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 ADCP Reporter Bioassay (THP-1) Shows Precision, Accuracy and Linearity.
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 100.9 | |
| 70 | 98.7 | |
| 100 | 101.4 | |
| 130 | 97.2 | |
| 150 | 97.9 | |
| Repeatability (% CV) | 100% (Reference) | 2.3 |
| Intermediate Precision (% CV) | 9.8 | |
| Linearity (r2) | 0.999 | |
| Linearity (y = mx + b) | y = 0.961x – 2.685 | |
|
A 50–150% theoretical potency series of rituximab (Anti-CD20) was analyzed in triplicate in three independent experiments performed on three days by two analysts using the ADCP Reporter Bioassay (THP-1) with Raji target cells. Bio-Glo-NL™ Reagent was added and luminescence quantified using the GloMax® Discover System. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells. |
||
Representative data for the ADCP Bioassay (THP-1). Panel A. ADCP Reporter Bioassay Effector Cells (THP-1) were incubated with serial dilutions of Control Ab, Anti-CD20, or trastuzumab (Anti-HER2) in the presence of Raji target cells (CD20+/HER2−). Panel B. ADCP Reporter Bioassay Effector Cells (THP-1) were incubated with serial dilutions of Control Ab, Anti-CD20, in the presence or absence of Raji target cells. After a 4-hour incubation at 37°C, 5% CO2, Bio-Glo-NL™ Reagent was added and luminescence was quantified using the GloMax® Discover System. Data were fitted to a four-parameter logistic curve using GraphPad Prism® software. Data were generated using thaw-and-use cells.
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
ADCP Bioassay Effector Cells (THP-1) |
GA128A | 1 × 0.5ml |
RPMI 1640 Medium |
G708A | 1 × 36ml |
Low IgG Serum |
G711A | 1 × 4ml |
Bio-Glo-NL™ Luciferase Assay Buffer |
J308A | 1 × 10ml |
Bio-Glo-NL™ Luciferase Assay Substrate |
J309A | 1 × 200μl |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.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-NL™ 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.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
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 |
|---|---|---|
ADCP Bioassay Effector Cells (THP-1) |
GA128A | 5 × 0.5ml |
RPMI 1640 Medium |
G708A | 5 × 36ml |
Low IgG Serum |
G711A | 5 × 4ml |
Bio-Glo-NL™ Luciferase Assay Buffer |
J308A | 5 × 10ml |
Bio-Glo-NL™ Luciferase Assay Substrate |
J309A | 5 × 200μl |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.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-NL™ 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.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
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 |
|---|---|---|
ADCP Bioassay Effector Cells (THP-1) (CPM) |
GA127A | 2 × 1ml |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.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-NL™ 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.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
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 |
|---|---|---|
ADCP Bioassay Effector Cells (THP-1) (CPM) |
GA127A | 50 × 1ml |
SDS
Search for SDS分析証明書
使用制限
For Research Use Only. Not for Use in Diagnostic Procedures.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-NL™ 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.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
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.
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