TIGIT/CD155 Blockade Bioassay
Measure Potency and Stability of Biologics Designed to Block TIGIT/CD155 Interaction
- 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:
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Catalog Number: J2201
Catalog Number: J2205
Catalog Number: J2092
Catalog Number: J2051
Catalog Number: J1921
Measure the ability of biologics to block immune checkpoint signals
TIGIT, also known as WUCAM and Vstm3, is an immune checkpoint protein expressed on lymphocytes. Highest expression levels are observed on effector CD4+ and CD8+ T cells, regulatory T cells and NK cells. TIGIT has several distinct mechanisms of action that inhibit lymphocyte activation. First, TIGIT is an inhibitory counterpart of the co-stimulatory receptor CD226. When TIGIT is present on the surface of lymphocytes, it outcompetes CD226 for CD155 binding and thus negates CD226 signaling. Second, TIGIT inhibits CD226 homodimerization in cis, preventing CD226 signaling. Third, the cytoplasmic tail of TIGIT contains an immunoreceptor tyrosine-based inhibitory motif (ITIM), which could potentially lead to inhibitory signaling.
The TIGIT/CD155 Blockade Bioassay reflects the mechanism of action (MOA) of biologics designed to block the TIGIT/CD155 interaction.
Control Ab, Anti-TIGIT, is a mouse monoclonal antibody that shows blocking activity of TIGIT with its ligand, CD155. It is intended for use as a control with the PD-1+TIGIT Combination Bioassay and the TIGIT/CD155 Blockade Bioassay. These assays can be used to measure the potency and stability of antibodies and other biologics designed to block the TIGIT/CD155 interaction.
TIGIT Negative Cells are for use as a negative control in the TIGIT/CD155 Blockade Bioassay. TIGIT Negative Cells are activated by engagement of the TCR/CD3 complex and costimulatory receptor CD28 co-engagement.
Assay Components
The assay consists of two genetically engineered cell lines:
TIGIT Effector Cells: Jurkat T cells expressing human TIGIT with a luciferase reporter driven by a native promoter that can respond to both TCR activation and CD226 co-stimulation
CD155 aAPC/CHO-K1 Cells: CHO-K1 cells engineered to express human CD155 with an engineered cell-surface protein designed to activate the TCR complex in an antigen-independent manner
When the two cell types are co-cultured, TIGIT inhibits CD226 activation and promoter-mediated luminescence. Addition of an anti-TIGIT antibody blocks the interaction of TIGIT with CD155 or inhibits the ability of TIGIT to prevent CD226 homodimerization, resulting in promoter-mediated luminescence.
The TIGIT/CD155 Blockade Bioassay is also available in CPM format, containing cryopreserved cells that can be thawed, propagated and banked for long-term use.
The Workflow
Traditional Method
'Cells as reagents' Method
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 TIGIT/CD155 Blockade Bioassay is a bioluminescent cell-based assay that overcomes the limitations of existing assays and can be used to measure the potency and stability of antibodies and other biologics targeting TIGIT.
Representation of the TIGIT/CD155 Blockade Bioassay. The bioassay consists of two genetically engineered cell lines, TIGIT Effector Cells and CD155 aAPC/CHO-K1 Cells. Panel A. When co-cultured, TIGIT inhibits CD226 pathway-activated luminescence. The addition of anti-TIGIT antibody blocks the TIGIT/CD155 interaction, thereby re-establishing CD226 pathway-activated luminescence, which can be detected in a dose-dependent manner by addition of BIo-Glo™ Reagent and quantitation with a luminometer. Panel B. When co-cultured with non-TIGIT-expressing Effector Cells, CD155 induces luminescence by activation of the CD226 pathway.
Assay Performance Data
Accuracy and Precision are critical to bioassays and the development of biologics. The TIGIT/CD155 Blockade Bioassay is 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. 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 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.
The TIGIT/CD155 Blockade Bioassay Shows Precision, Accuracy and Linearity.
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 101.8 | |
| 70 | 102.8 | |
| 140 | 101.0 | |
| 200 | 98.5 | |
| Repeatability (% CV) | 100% (Reference) | 11.0 |
| Intermediate Precision (% CV) | 9.7 | |
| Linearity (r2) | 0.999 | |
| Linearity (y = mx + b) | y = 0.974x + 3.33 | |
| A 50–200% theoretical potency series of Control Ab, Anti-TIGIT, was analyzed in triplicate in three independent experiments performed on three days by two analysts. Bio-Glo™ Reagent was added and luminescence quantified using the GloMax® Discover Detection System. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells. | ||
The TIGIT/CD155 Blockade Bioassay reflects the mechanism of action (MOA) and specificity of biologics designed to block the TIGIT/CD155 interaction.
The TIGIT/CD155 Blockade Bioassay measures the activity of Control Ab, Anti-TIGIT.
Product Kit Formats and Related Products
- TIGIT/CD155 Blockade Bioassay is available in 1X and 5X sizes.
- TIGIT/CD155 Blockade Bioassay is also available in CPM format (Cat.# J2092).
- Control Ab, Anti-TIGIT, is available separately (Cat.# J2051).
- TIGIT Negative Cells are available separately (Cat.# J1921).
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
TIGIT Effector Cells |
J169A | 1 × 0.5ml |
CD155 aAPC/CHO-K1 Cells |
J180A | 1 × 0.5ml |
Fetal Bovine Serum |
J121A | 1 × 4ml |
RPMI 1640 Medium |
G708A | 1 × 36ml |
Bio-Glo™ Luciferase Assay Buffer |
G719A | 2 × 10ml |
Bio-Glo™ Luciferase Assay Substrate |
G720A | 2 × 1 vial |
SDS
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Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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.
What's in the box?
| Item | Part # | Size |
|---|---|---|
TIGIT Effector Cells |
J169A | 5 × 0.5ml |
CD155 aAPC/CHO-K1 Cells |
J180A | 5 × 0.5ml |
Fetal Bovine Serum |
J121A | 5 × 4ml |
RPMI 1640 Medium |
G708A | 5 × 36ml |
Bio-Glo™ Luciferase Assay Buffer |
G719A | 10 × 10ml |
Bio-Glo™ Luciferase Assay Substrate |
G720A | 10 × 1 vial |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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.
What's in the box?
| Item | Part # | Size |
|---|---|---|
TIGIT Effector Cells (CPM) |
J168A | 2 × 1ml |
CD155 aAPC/CHO-K1 Cells (CPM) |
J179A | 2 × 1ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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.
What's in the box?
| Item | Part # | Size | Concentration |
|---|---|---|---|
Control Ab, Anti-TIGIT |
J205A | 1 × 100μg | 1mg/ml |
SDS
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Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
Please see the Protocol for detailed storage conditions.
The transfer of this product is conditioned on the buyer using the purchased product for (i) research use, (ii) discovery, development and monitoring of biologic drugs and vaccines, (iii) quality assurance and quality control testing of biologic drugs and vaccines, and (iv) product release assays for biologic drugs and vaccines conducted by the buyer, including contract research or any fee for service research, and the buyer must not (1) use this product or its components for (1a) any use, indirectly (except as allowed in (iii) and (iv) above) or directly in manufacturing, or (1b) diagnostic, therapeutic or prophylactic purposes; and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research.
What's in the box?
| Item | Part # | Size |
|---|---|---|
TCR/CD3 Effector Cells (IL-2) |
J129A | 1 × 0.8ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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.
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