SIRPα/CD47 Blockade Bioassay, Fc-Dependent

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Measure Potency and Stability of Biologics that Bind and Block SIRPα/CD47 Interactions

  • Stability-indicating 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

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Catalog number selected: JA4801

$ 1,420.00
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SIRPα/CD47 Blockade Bioassay, Fc-Dependent
Thaw and Use/1X each component
$ 1,420.00
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Convenient Bioassay, Quality Results

Antibody-dependent cellular phagocytosis (ADCP) is an important mechanism-of-action (MOA) for antibody-based immunotherapies. ADCP is regulated by a complex network of checkpoints, including SIRPα and CD47, that facilitate removal of aberrant or infected cells while preserving healthy tissues. Despite its important physiological role, the SIRPα/CD47 checkpoint contributes to immune escape by tumor cells, many of which overexpress CD47 and thereby evade phagocytosis. Biologics that inhibit SIRPα/CD47 interaction enhance phagocytosis of tumor cells in vitro and have shown promising therapeutic efficacy.

Current methods for assessing the activity of SIRPα/CD47 checkpoint inhibitors rely on primary monocyte-derived 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 a quality-controlled setting.

The SIRPα/CD47 Blockade Bioassay, Fc-Dependent, is a bioluminescent reporter cell-based assay used to measure the potency and stability of ligands or antibodies that bind and block SIRPα and CD47 interactions.

Bioassay Components and Product Formats

The blockade bioassay consists of two genetically engineered cell lines:

SIRPα Effector Cells: A monocytic cell line with a stably integrated NanoLuc® (NL) luciferase reporter driven by FcγR and SIRPα/CD47 pathway-dependent response elements.

CD47 CHO-K1 Target Cells: CHO-K1 cells engineered to express human CD47. These cells do not provide an activating signal for FcγRs.

Thaw-and-Use Kits (1X, 5X):

  • Include SIRPα Effector Cells, CD47 CHO-K1 Target Cells, cell culture media and assay reagents
  • Recommended for routine use with customer-defined antibody
  • Available in 1X and 5X sizes

Cell Propagation Model format (CPM; Cat.# JA4802):

  • Includes the SIRPα Effector Cells and CD47 CHO-K1 Target Cells (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.# GA6000, GA6030):

  • 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 the two cell types are co-cultured, SIRPα/CD47 interaction inhibits FcγR signaling and promoter-driven luminescence. Addition of Fc-functional anti-SIRPα or anti-CD47 antibody that blocks SIRPα/CD47 interaction releases the inhibitory signal and results in FcγR activation and promoter-driven luminescence.

Workflow

Standard bioassay workflow with cultured cells
Improved bioassay workflow with thaw-and-use cells

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 SIRPα/CD47 Blockade Bioassay, Fc-Dependent, measures the potency and stability of biologics designed to block the SIRPα/CD47 interaction and activate FcγRs.

Representation of the SIRPα/CD47 Blockade Bioassay, Fc-Dependent.

Representation of the SIRPα/CD47 Blockade Bioassay, Fc-Dependent. The SIRPα/CD47 Blockade Bioassay, Fc-Dependent, consists of SIRPα Effector Cells, CD47 CHO-K1 Target Cells and an Fc-functional SIRPα/CD47 inhibitor. When all components are co-cultured, the Fc-functional SIRPα/CD47 inhibitor disrupts the SIRPα/CD47 interaction and activates FcγRs on the surface of the effector cells. This results in promoter-driven luminescence that can be detected and quantified using Bio-Glo-NL™ Reagent.

Assay Performance Data

Representative data for the SIRPα/CD47 Blockade Bioassay, Fc-Dependent (Panel A).
Representative data for the SIRPα/CD47 Blockade Bioassay, Fc-Dependent (Panel B).

Representative data for the SIRPα/CD47 Blockade Bioassay, Fc-Dependent. Panel A. SIRPα Effector Cells were incubated with CD47 CHO-K1 Target Cells in the presence of serial titrations of blocking antibodies as indicated. Panel B. The SIRPα Effector Cells were incubated with CD47 CHO-K1 Target Cells in the presence of anti-CD47 blocking [B6.H12] or non-blocking [2D3] antibody. After a 4-hour induction, Bio-Glo-NL™ Reagent was added and luminescence 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

You are viewing: JA4801 Change Configuration

What's in the box?

Item Part # Size

SIRPα Effector Cells

GA131A 1 × 0.5ml

CD47 CHO-K1 Target Cells

GA126A 1 × 0.5ml

Fetal Bovine Serum

J121A 1 × 4ml

Ham's F-12 Medium

J123A 1 × 25ml

RPMI 1640 Medium

G708A 1 × 36ml

Bio-Glo-NL™ Luciferase Assay Buffer

J308A 1 × 10ml

Bio-Glo-NL™ Luciferase Assay Substrate

J309A 1 × 200μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

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. 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.

U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.

Product cannot be used for proficiency testing.

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.

Specifications

You are viewing: JA4805 Change Configuration

What's in the box?

Item Part # Size

SIRPα Effector Cells

GA131A 5 × 0.5ml

CD47 CHO-K1 Target Cells

GA126A 5 × 0.5ml

Fetal Bovine Serum

J121A 5 × 4ml

Ham's F-12 Medium

J123A 5 × 25ml

RPMI 1640 Medium

G708A 5 × 36ml

Bio-Glo-NL™ Luciferase Assay Buffer

J308A 5 × 10ml

Bio-Glo-NL™ Luciferase Assay Substrate

J309A 5 × 200μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

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.

Product cannot be used for proficiency testing.

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.

Specifications

You are viewing: JA4802 Change Configuration

What's in the box?

Item Part # Size

CD47 CHO-K1 Target Cells (CPM)

GA124A 2 × 1ml

SIRPα Effector Cells (CPM)

GA129A 2 × 1ml

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

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. 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.

Product cannot be used for proficiency testing.

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.

Specifications

You are viewing: GA6000 Change Configuration

What's in the box?

Item Part # Size

SIRPα Effector Cells (CPM)

GA129A 50 × 1ml

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

XX

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

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.

Product cannot be used for proficiency testing.

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.

Specifications

You are viewing: GA6030 Change Configuration

What's in the box?

Item Part # Size

CD47 CHO-K1 Target Cells (CPM)

GA124A 50 × 1ml

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

XX

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

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.

Product cannot be used for proficiency testing.

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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