PD-1+TIGIT Combination Bioassay
The assay consists of two genetically engineered cell lines:
PD-1+TIGIT Effector Cells: Jurkat T cells engineered to express human PD-1 and human TIGIT with a luciferase reporter driven by a native promoter that can respond to both TCR activation and CD226 co-stimulation
PD-L1+CD155 aAPC/CHO-K1 Cells: CHO-K1 cells engineered to express human PD-L1 and 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,PD-1 binds to PD-L1 and inhibits TCR-induced activation and promoter-mediated luminescence. Addition of an anti-PD-1 Ab or an anti-TIGIT Ab blocks the interaction of PD-1 with PD-L1 or TIGIT with CD155, respectively, and results in a modest enhancement in promoter-mediated luminescence. Addition of antibodies that block both PD-1/PD-L1 and TIGIT/CD155 will result in much higher promoter-mediated luminescence due to synergy between these pathways.
'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 PD-1+TIGIT Combination 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 PD-1 and TIGIT.
Assay Performance Data
Accuracy and Precision are critical to bioassays and the development of biologics. The PD-1+TIGIT Combination 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.
Measure of ability to replicated a reference or "target" value.
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).
Measure of repeatability of multiple assays of the same homogeneous sample with different analysts or equipment or on different days.
Ability of your assay to provide measured values that are proportional to the concentration of the sample.
Expected Relative Potency Recovery 50% 95.3% 70% 105.5% 140% 104.9% 200% 109.9%
Repeatability (100% reference, %CV)
Intermediate Precision (%CV)
r 2 = 0.999 (50-200% range)
y = 1.131x + 0.028
The PD-1+TIGIT Combination Bioassay reflects the mechanism of action (MOA) and specificity of biologics designed to block the PD-1/PD-L1 and TIGIT/CD155 interaction.
The PD-1+TIGIT Combination Bioassay measures the activity of Control Ab, Anti-PD-1, and Control Ab, Anti-TIGIT.
PD-1+TIGIT Combination Bioassay Technical Manual
PDF (911 KB)
SpecificationsWhat's in the box?
Item Part # Size
PD-1+TIGIT Effector Cells
J176A 1 × 1 vial
PD-L1+CD155 aAPC/CHO-K1 Cells
J184A 1 × 1 vial
Fetal Bovine Serum
J121A 1 × 4ml
Bio-Glo™ Luciferase Assay Buffer
G719A 1 × 10ml
Bio-Glo™ Luciferase Assay Substrate
G720A 1 × 1 vial
Ham's F-12 Medium
J123A 1 × 25ml
RPMI 1640 Medium
G708A 1 × 36ml
Use RestrictionsNot For Medical Diagnostic Use.
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 and European Pat. No. 1341808.
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.