human pd1 fc protein  (R&D Systems)

Journal: Journal of Cellular and Molecular Medicine

Article Title: FoxO3a‐Mediated Modulation of PD‐L1 Expression and Inhibition by Dihydroartemisinin in Triple‐Negative Breast Cancer

doi: 10.1111/jcmm.70947

Figure Lengend Snippet: DA treatment sensitises MDA‐MB‐231 cells to activated T cells. Representative images (A) and quantification (B) of green fluorescence‐labelled PD1/Fc on DA‐treated MDA‐MB‐231 cells for 24 h. Representative images (C) and quantification (D) of crystal violet‐stained live cancer cells in T cell‐mediated cancer cell killing assay. MDA‐MB‐231 cells were then co‐cultured with activated T cells for 48 h with or without DA (12.5 μM) and subjected to crystal violet staining. Statistical significance was determined using * p < 0.05 and ** p < 0.01. All error bars are expressed as mean ± SD of three independent experiments. Abbreviations: DA, dihydroartemisinin; CTRL, negative control.

Article Snippet: Cells in each group were fixed in 4% PFA for 15 min, incubated with recombinant human PD1 Fc protein (R&D Systems, Minneapolis, MN, USA) for 1 h, and then incubated with anti‐human Alexa Fluor 488 secondary antibodies (Thermal Fisher Scientific) for 1 h at room temperature.

Techniques: Fluorescence, Staining, Cell Culture, Negative Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Design and selection of anti-PD-L1 single-domain antibody and tumor necrosis factor superfamily ligands for an optimal vectorization in an oncolytic virus

doi: 10.3389/fbioe.2023.1247802

Figure Lengend Snippet: Single-chain dimeric anti-hPDL1 sdAb GS542 is the best vaccinia virus vectorized constructs for blocking PD1/PDL1 interactions. Purified sdAb (also mentioned as GS542) or avelumab (A, B) or the culture medium of A549 or MIA PaCa2 cells infected by the indicated recombinant virus (i.e., VV-xx: (C–E) were analyzed for transgene expression by immunoblot using an HRP-conjugated anti-6xHis tag monoclonal antibody and (C) competition of PD1/PDL1 interaction either by ELISA (A, D) or using cellular bioassay (B, E) . Inserted graph in represents the calculated effective concentration 50% (EC50) +/- confidence interval (CI) 95% for each curve. Mock: culture medium of cells infected with a VV encoding no transgene; scDim: VV encoding single-chain dimeric sdAb GS542; Mono: VV encoding monomeric sdAb GS542; Avel: VV encoding avelumab; Dim. Hinge: VV encoding GS542 fused to IgG2 hinge; and Fc fusion: VV encoding sdAb fused to human IgG1 Fc.

Article Snippet: Human PD1 Fc (R&D systems, 1086-PD) was coated on the 96-well ELISA plate at 0.25 μg/mL in 50 mM carbonate buffer at pH 9.6.

Techniques: Virus, Construct, Blocking Assay, Purification, Infection, Recombinant, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Bioassay, Concentration Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Design and selection of anti-PD-L1 single-domain antibody and tumor necrosis factor superfamily ligands for an optimal vectorization in an oncolytic virus

doi: 10.3389/fbioe.2023.1247802

Figure Lengend Snippet: The CD40 agonist activity of sdAb–linker–CD40L is inhibited by coincubation with a PDL1 blocking antibody, and sdAb–linker–CD40L has a PD1/PDL1 blocking activity. CD40 agonist activity of culture supernatants of VV–sdAb–linker–CD40L-infected cells was analyzed, as shown in , in the presence or absence of 100 μg/mL of PDL1 blocking antibody (avelumab). In the case of the addition of avelumab, or of its isotype control, only undiluted supernatants were tested (A) . The PD1/PDL1 blocking activity of the culture medium of A549 cells infected by the mentioned virus was measured, as shown in (B) . VV–sdAb–link–CD40L is a vaccinia virus encoding the sdAb–link–CD40L construct. VV–scDimeric: VV encoding single-chain dimeric sdAb GS542; VV–monomeric: VV encoding monomeric sdAb GS542; and VV–avelumab: VV encoding avelumab.

Article Snippet: Human PD1 Fc (R&D systems, 1086-PD) was coated on the 96-well ELISA plate at 0.25 μg/mL in 50 mM carbonate buffer at pH 9.6.

Techniques: Activity Assay, Blocking Assay, Infection, Control, Virus, Construct