TIGIT/CD155 Blockade Bioassay

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

Standard bioassay workflow with cultured cells

'Cells as reagents' Method

Improved bioassay workflow with thaw-and-use cells

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.

14857ma-w — **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.

14858ma-w — TIGIT Effector Cells were incubated with CD155 aAPC/CHO-K1 Cells in the presence of a serial titration of Control Ab, Anti-TIGIT (Cat.# J2051), anti-PD-1 (nivolumab), anti-CTLA-4 (ipilimumab) or anti-PD-L1 blocking Abs as indicated.

14923ma-w — TIGIT Effector Cells were plated overnight. The following day, a titration fo Control Ab, Anti-TIGIT, was added followed by CD155 aAPC/CHO-K1 Cells.

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).

Protocolos

Complete Protocol

TIGIT/CD155 Blockade Bioassay Technical ManualPDF (2166 KB) – English

TIGIT/CD155 Blockade Bioassay, Propagation Model Technical ManualPDF (1037 KB) – English

Contenido

Item	Part #	Presentación
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

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Condiciones de Almacenaje

Please see the Protocol for detailed storage conditions.

Patentes y Exclusiones

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.

U.S. Pat. No. 8,008,006.

Patent Pending.

U.S. Pat. No. 10,077,244 and other patents.

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.

Product cannot be used for proficiency testing.

Contenido

Item	Part #	Presentación
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 SDS

Certificado de Análisis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Condiciones de Almacenaje

Please see the Protocol for detailed storage conditions.

Patentes y Exclusiones

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.

Patent Pending.

U.S. Pat. No. 8,008,006.

U.S. Pat. No. 10,077,244 and other patents.

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.

Contenido

Item	Part #	Presentación
TIGIT Effector Cells (CPM)	J168A	2 × 1ml
CD155 aAPC/CHO-K1 Cells (CPM)	J179A	2 × 1ml

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Condiciones de Almacenaje

Please see the Protocol for detailed storage conditions.

Patentes y Exclusiones

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.

Patent Pending.

U.S. Pat. No. 8,008,006.

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.

Contenido

Item	Part #	Presentación	Concentración
Control Ab, Anti-TIGIT	J205A	1 × 100μg	1mg/ml

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Condiciones de Almacenaje

Please see the Protocol for detailed storage conditions.

Patentes y Exclusiones

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.

Contenido

Item	Part #	Presentación
TCR/CD3 Effector Cells (IL-2)	J129A	1 × 0.8ml

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Condiciones de Almacenaje

Please see the Protocol for detailed storage conditions.

Patentes y Exclusiones

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.

U.S. Pat. No. 8,008,006.

Recursos

Posters

Otros

Immune Checkpoint Bioassays Power Combination Immunotherapy

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TIGIT/CD155 Blockade Bioassay

Measure Potency and Stability of Biologics Designed to Block TIGIT/CD155 Interaction

Measure the ability of biologics to block immune checkpoint signals

Assay Components

The Workflow

The Assay Principle

Assay Performance Data

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

Protocolos

Complete Protocol

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Recursos

Posters

Otros

Productos Similares

PD-1+TIGIT Combination Bioassay

Usado con frecuencia junto con

GloMax® Discover System

Bio-Glo™ Luciferase Assay System

Americas

Pacific Asia

Europe