pd-1 jurkat reporter cells promega, cs187102  (Promega)

Journal: mAbs

Article Title: Engineering a tumor-selective prodrug T-cell engager bispecific antibody for safer immunotherapy

doi: 10.1080/19420862.2024.2373325

Figure Lengend Snippet: Preclinical functional validation of pro-TCE with recombinant MMP-2 protease treatment. All experiments were performed at an effector cell to target cell ratio of 1:1. Each point represents the mean value of triplicates. (a) T cell activation induced by pro-TCE after proteolysis using a Jurkat NFAT-Luciferase T cell model. (b) HER2 + target cell killing by hPBMCs induced by pro-TCE after proteolysis after 48 hours. (c–d) IL-2 and IFN-γ secretion by hPBMCs induced by pro-TCE after 48 hours. The reported errors for the EC 50 values are the 95% confidence intervals from fitting each individual experiment.

Article Snippet: Jurkat NFAT-Luciferase reporter cells (Promega, J1601) and primary human T cells (ALLCELLS frozen normal human peripheral blood CD3 + Pan T cells, include cat#260100.19) were grown in RPMI growth media containing 10% fetal bovine serum and hygromycin B (Gibco, #10687–010).

Techniques: Functional Assay, Biomarker Discovery, Recombinant, Activation Assay, Luciferase

Journal: mAbs

Article Title: Engineering a tumor-selective prodrug T-cell engager bispecific antibody for safer immunotherapy

doi: 10.1080/19420862.2024.2373325

Figure Lengend Snippet: NCI-N87 tumor cell secreted proteases trigger pro-TCE activity. SDS-PAGE gel electrophoresis of pro-TCE before and after incubation with NCI-N87 cellular supernatant for 1 h (left). Jurkat NFAT-Luciferase T cell activation assay with NCI-N87 target cells (right). The reported EC 50 value is an average of three replicates with errors reflecting the 95% confidence intervals of the average measurements.

Article Snippet: Jurkat NFAT-Luciferase reporter cells (Promega, J1601) and primary human T cells (ALLCELLS frozen normal human peripheral blood CD3 + Pan T cells, include cat#260100.19) were grown in RPMI growth media containing 10% fetal bovine serum and hygromycin B (Gibco, #10687–010).

Techniques: Activity Assay, SDS Page, Nucleic Acid Electrophoresis, Incubation, Luciferase, Activation Assay

Journal: mAbs

Article Title: Engineering a tumor-selective prodrug T-cell engager bispecific antibody for safer immunotherapy

doi: 10.1080/19420862.2024.2373325

Figure Lengend Snippet: Expanding the pro-TCE conditionally active T cell engager platform to clone SP34. (a) Anti-CD3 mAbs hE10 and SP34 both bind the N-terminus of CD3ε. (b) SDS-PAGE gel electrophoresis of SP34_Pertuzumab pro-TCE before and after proteolysis. (c) Designed pro-TCE using anti-CD3 mAb with SP34 CD3-binding arm activates Jurkat NFAT-Luciferase T cells when proteolyzed but not when masked. The reported EC 50 value is an average of three replicates with errors reflecting the 95% confidence intervals of the average measurements.

Article Snippet: Jurkat NFAT-Luciferase reporter cells (Promega, J1601) and primary human T cells (ALLCELLS frozen normal human peripheral blood CD3 + Pan T cells, include cat#260100.19) were grown in RPMI growth media containing 10% fetal bovine serum and hygromycin B (Gibco, #10687–010).

Techniques: SDS Page, Nucleic Acid Electrophoresis, Binding Assay, Luciferase