Cat. No：GM-032AS004
Product：H_PD-1 PDL1 Reporter Blockade Assay (Raji)
Cat. No：GM-032AS004
Product：H_PD-1 PDL1 Reporter Blockade Assay (Raji)
H_PD-1 Reporter Jurkat Cell Line
Cell Growth Medium：RPMI 1640+10% FBS+1% P.S+3.5 μg/mL Blasticidin+0.75 μg/mL Puromycin
Cell Freezing Medium：90% FBS+10% DMSO
Raji Cell Line
Cell Growth Medium：RPMI 1640+10% FBS+1% P.S
Cell Freezing Medium：90% FBS+10% DMSO
Assay Buffer：RPMI 1640+1% FBS+1% P.S
PD-1 is an immune inhibitory receptor expressed on activated T cells and B cells, playing a crucial role in regulating immune responses to tumor antigens and self-antigens. The interaction between PD-1 on neighboring cells and its ligands, PD-L1 or PD-L2, inhibits the transmission of the TCR signaling pathway, as well as TCR-mediated cell proliferation, transcription activation, and cytokine production. Therapeutic antibodies and Fc fusion proteins that block the PD-1/PD-L1 interaction have shown promising prospects in clinical trials for treating various cancers.
Current methods for evaluating the activity of anti-PD-1 or anti-PD-L1 biologics rely on the measurement of primary human T cells and functional endpoints, such as cell proliferation, cell surface marker expression, interferon γ (IFNγ), and interleukin-2 (IL-2) production. These assays are labor-intensive and subject to variability due to reliance on donor primary cells, complex experimental procedures, and inadequate assay reagents, making it challenging to establish these assays in a quality-controlled drug development environment.
The H_PD-1 PDL1 Reporter Blockade Assay (Raji) is a biologically relevant mechanism of action-based assay that can be used to assess the efficacy and stability of antibodies and other biologics that block the PD-1/PD-L1 interaction. This assay utilizes two genetically engineered cell lines: PD-1 Effector Cells, stable Jurkat T cells expressing human PD-1 receptor and NFAT-induced luciferase reporter gene; and Raji H_PDL1 cell line, a stable Raji cell line expressing human PD-L1 and a cell surface protein that activates a homologous TCR in an antigen-independent manner.
When these two cell lines are co-cultured, the interaction between PD-1 and PD-L1 inhibits the transmission of the TCR signaling pathway and NFAT-mediated luciferase expression. Addition of antibodies blocking PD-1/PD-L1 reverses this inhibition, resulting in the transmission of the TCR signaling pathway and NFAT-mediated luciferase expression.
Cat. No：GM-032AS004
Product：H_PD-1 PDL1 Reporter Blockade Assay (Raji)
H_PD-1 Reporter Jurkat Cell Line
Cell Growth Medium：RPMI 1640+10% FBS+1% P.S+3.5 μg/mL Blasticidin+0.75 μg/mL Puromycin
Cell Freezing Medium：90% FBS+10% DMSO
Raji Cell Line
Cell Growth Medium：RPMI 1640+10% FBS+1% P.S
Cell Freezing Medium：90% FBS+10% DMSO
Assay Buffer：RPMI 1640+1% FBS+1% P.S
PD-1 is an immune inhibitory receptor expressed on activated T cells and B cells, playing a crucial role in regulating immune responses to tumor antigens and self-antigens. The interaction between PD-1 on neighboring cells and its ligands, PD-L1 or PD-L2, inhibits the transmission of the TCR signaling pathway, as well as TCR-mediated cell proliferation, transcription activation, and cytokine production. Therapeutic antibodies and Fc fusion proteins that block the PD-1/PD-L1 interaction have shown promising prospects in clinical trials for treating various cancers.
Current methods for evaluating the activity of anti-PD-1 or anti-PD-L1 biologics rely on the measurement of primary human T cells and functional endpoints, such as cell proliferation, cell surface marker expression, interferon γ (IFNγ), and interleukin-2 (IL-2) production. These assays are labor-intensive and subject to variability due to reliance on donor primary cells, complex experimental procedures, and inadequate assay reagents, making it challenging to establish these assays in a quality-controlled drug development environment.
The H_PD-1 PDL1 Reporter Blockade Assay (Raji) is a biologically relevant mechanism of action-based assay that can be used to assess the efficacy and stability of antibodies and other biologics that block the PD-1/PD-L1 interaction. This assay utilizes two genetically engineered cell lines: PD-1 Effector Cells, stable Jurkat T cells expressing human PD-1 receptor and NFAT-induced luciferase reporter gene; and Raji H_PDL1 cell line, a stable Raji cell line expressing human PD-L1 and a cell surface protein that activates a homologous TCR in an antigen-independent manner.
When these two cell lines are co-cultured, the interaction between PD-1 and PD-L1 inhibits the transmission of the TCR signaling pathway and NFAT-mediated luciferase expression. Addition of antibodies blocking PD-1/PD-L1 reverses this inhibition, resulting in the transmission of the TCR signaling pathway and NFAT-mediated luciferase expression.
