PD-1/PD-L1 Blockade Bioassay Protocol
Instructions for Use of Product(s)
J1250, J1255, J3011, J3015, J4011, J4015, J1191, J1195
Literature # TM468
The PD-1/PD-L1 Blockade Bioassay is a bioluminescent cell-based assay that can be used to measure the potency and stability of antibodies and other biologics designed to block the PD-1/PD-L1 interaction. The assay consists of two genetically engineered cell lines: PD-1 Effector Cells, which are Jurkat T cells expressing human PD-1 and a luciferase reporter driven by an NFAT response element (NFAT-RE), and; PD-L1 aAPC/CHO-K1 Cells, which are CHO-K1 cells expressing human PD-L1 and an engineered cell surface protein designed to activate cognate TCRs in an antigen-independent manner.
The PD-1 Effector Cells and PD-L1 aAPC/CHO-K1 Cells are provided in thaw-and-use format as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell propagation. When the two cell types are co-cultured, the PD-1/PD-L1 interaction inhibits TCR signaling and NFAT-RE-mediated luminescence. Addition of either an anti-PD-1 or anti-PD-L1 antibody that blocks the PD-1/PD-L1 interaction releases the inhibitory signal and results in TCR activation and NFAT-RE-mediated luminescence. The PD-1/PD-L1 Blockade Bioassay includes the necessary medium and serum to thaw, plate and assay the cells. The bioluminescent signal can be detected and quantified using the Bio-Glo™ Luciferase Assay System, also included in the kit, and a standard luminometer such as the GloMax® Discover System.
This Technical Manual is relevant for Cat.# J1250, J1255, J1191 and J1195; J3011 and J3015 (Taiwan); J4011 and J4015 (Korea).
Summary of Changes
The following changes were made to the 10/21 revision of this document:
1. A new Limited Use statement was added on pages 2 and 23.
2. The equation in 4.G was reformatted.
3. Section 7.B, Related Products, was updated.
4. The cover image was updated
Revised 10/21.