In cell-based assays, REGN2810 reverses PD-1-dependent attenuation of T-cell receptor signaling in engineered T cells and enhances responses of human primary T cells.
Cemiplimab (REGN2810) binds the humanized PD-1 receptor and inhibits growth of MC38 murine tumors in human PD-1 knock-in mice. As REGN2810 binds to cynomolgus monkey PD-1 with high affinity, pharmacokinetic and toxicologic assessment of REGN2810 is performed in cynomolgus monkeys. High doses of REGN2810 are well tolerated, without adverse immune-related effects.
Objective: T cell activation bioassays Cells: Jurkat/AP-1-Luc/hPD-1 cells, HEK293/hCD20 cells, HEK293/hCD20/hPD-L1 cells Concentrations: 0.1 pM-0.1 μM Incubation Time: 4-6 h Method: Antigenpresenting cell (APC)-like HEK293 cells are generated by lentiviral transduction of human CD20 and human PD-L1. T-cell receptor (TCR) activation is achieved by an antiCD3 x anti-CD20 bispecific antibody. To generate a dose response curve for anti-CD3 x anti-CD20, the bispecific molecule is serially diluted and tested with 50,000/well Jurkat/AP-1-Luc/hPD-1 and 10,000/well HEK293/hCD20 or HEK293/hCD20/hPD-L1 cells in a 96-well plate. Serially diluted REGN2810 is tested under similar conditions in the presence of a fixed concentration of anti-CD3 x anti-CD20 (100 pM). Plates are incubated at 37°C for 4-6 hours. Reference: https://pubmed.ncbi.nlm.nih.gov/28265006/
Objective: T cell activation bioassays Cells: HEK293/hCD20/hPD-L1 cells, CD4+ T cells, HEK293/hCD20/hPD-L1 cells Concentrations: 0.1 pM-0.1 μM Incubation Time: 72 h Method: HEK293/hCD20/hPD-L1 cells are treated with 50 mg/mL mitomycin C for 30 minutes at 37°C to inhibit proliferation. Serially diluted REGN2810 is incubated with 50,000/well preactivated CD4+ T cells and 25,000/well of HEK293/hCD20/hPD-L1 cells in the presence of 2 nM anti-CD3 x anti-CD20 bispecific antibody in a 96-well plate for 72 hours. Reference: https://pubmed.ncbi.nlm.nih.gov/28265006/
Cemiplimab can apply to nude mice, various cancer cell lines and other related assays (Only for Reference)