fc tagged human pd 1 protein  (Sino Biological)

Journal: Translational Oncology

Article Title: Peptide-based inhibitors targeting the PD-1/PD-L1 axis: potential immunotherapeutics for cancer

doi: 10.1016/j.tranon.2024.101892

Figure Lengend Snippet: Structure of the PD-1/PD-L1 complex (PDB: 4ZQK ) with the designed peptide groups marked on it; PD-1: blue, PD-L1: green. The PD-L1 fragments used to design three groups of peptides are marked as follows: purple - Group I, yellow - Group II, red - Group III. The amino acids in PD-L1 crucial for interaction with PD-1 are represented as sticks.

Article Snippet: Subsequently, 2-fold serial dilutions of PD-1-Fc (Sino Biological Company, China, #10084-H08H-B) in PBS-T, at concentrations from 8.00 to 0.5 µg/ml, were added to the precoated wells.

Techniques:

Journal: Translational Oncology

Article Title: Peptide-based inhibitors targeting the PD-1/PD-L1 axis: potential immunotherapeutics for cancer

doi: 10.1016/j.tranon.2024.101892

Figure Lengend Snippet: Binding of PD-1 to PD-L1 or to the indicated PD-L1-derived peptides analysed by indirect ELISA. Results are shown for at least three experiments performed independently in duplicate. Data are depicted as mean ± SD. Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by Dunnet's post-hoc test. ****: p < 0.0001, ***: p < 0.001, **: p < 0.01, *: p < 0.05.

Article Snippet: Subsequently, 2-fold serial dilutions of PD-1-Fc (Sino Biological Company, China, #10084-H08H-B) in PBS-T, at concentrations from 8.00 to 0.5 µg/ml, were added to the precoated wells.

Techniques: Binding Assay, Derivative Assay, Indirect ELISA

Journal: Translational Oncology

Article Title: Peptide-based inhibitors targeting the PD-1/PD-L1 axis: potential immunotherapeutics for cancer

doi: 10.1016/j.tranon.2024.101892

Figure Lengend Snippet: Sensorgrams of the PD-L1 derived peptides interacting with PD-1, obtained using the SPR technique. ND - not determined (no binding detected or binding too weak to establish reliable constants).

Article Snippet: Subsequently, 2-fold serial dilutions of PD-1-Fc (Sino Biological Company, China, #10084-H08H-B) in PBS-T, at concentrations from 8.00 to 0.5 µg/ml, were added to the precoated wells.

Techniques: Derivative Assay, Binding Assay

Journal: Translational Oncology

Article Title: Peptide-based inhibitors targeting the PD-1/PD-L1 axis: potential immunotherapeutics for cancer

doi: 10.1016/j.tranon.2024.101892

Figure Lengend Snippet: Competitive inhibition of PD-1/PD-L1 complex formation by the peptides obtained in this study. Modified Jurkat cells (JE6–1-NF-κB::eGFP PD-1) were incubated with the PD-L1-derived peptides, with the final concentration used in the experiment ranging from 50.00 to 5.56 µM. Subsequently, the cells were probed with PD-L1-Fc followed by PE-labelled anti-human IgG antibody and analysed by flow cytometry. The bar diagram shows the fold induction of geometric mean fluorescent intensity (gMFI) for at least three experiments performed independently in triplicate. Data were normalized to the gMFI obtained for the reporter PD-1 cell line treated with PD-L1-Fc in the absence of peptides. Data are depicted as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Dunnet's post-hoc test. ****: p < 0.0001, ***: p < 0.001, **: p < 0.01, *: p < 0.05.

Article Snippet: Subsequently, 2-fold serial dilutions of PD-1-Fc (Sino Biological Company, China, #10084-H08H-B) in PBS-T, at concentrations from 8.00 to 0.5 µg/ml, were added to the precoated wells.

Techniques: Inhibition, Modification, Incubation, Derivative Assay, Concentration Assay, Flow Cytometry

Journal: Translational Oncology

Article Title: Peptide-based inhibitors targeting the PD-1/PD-L1 axis: potential immunotherapeutics for cancer

doi: 10.1016/j.tranon.2024.101892

Figure Lengend Snippet: Inhibitory properties of the peptides in the functional cellular reporter assay. The PD-1 reporter cells were stimulated with TCS CD86/PD-L1 in the absence or presence of the peptides. The inhibitory properties of the peptides were measured based on eGFP expression by flow cytometry and normalized to gMFI eGFP obtained for the PD-1 reporter/TCS CD86 cells treated with the peptides. The dotted line shows the normalized eGFP expression level from the co-culture of the PD-1 reporter cells with TCS CD86/PD-L1. Results are shown for three experiments performed independently in duplicate. Data are depicted as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Dunnet's post-hoc test. ****: p < 0.0001, ***: p < 0.001, **: p < 0.01, *: p < 0.05.

Article Snippet: Subsequently, 2-fold serial dilutions of PD-1-Fc (Sino Biological Company, China, #10084-H08H-B) in PBS-T, at concentrations from 8.00 to 0.5 µg/ml, were added to the precoated wells.

Techniques: Functional Assay, Reporter Assay, Expressing, Flow Cytometry, Co-Culture Assay

Journal: Translational Oncology

Article Title: Peptide-based inhibitors targeting the PD-1/PD-L1 axis: potential immunotherapeutics for cancer

doi: 10.1016/j.tranon.2024.101892

Figure Lengend Snippet: Structures A and B show, respectively, the complexes between PD-1 (blue) and PD-L1 (green), and between PD-1 (blue) and the PD-L1(111–127) fragment (red) trimmed from the PD-1/PD-L1 complex (PDB code: 4ZQK). Structures C-F show the complexes of peptide ( L11 ) obtained from docking NMR structures to PD-1 by using the UNRES force field: (C) cluster from family 1 (lowest energy) – RMSD 3.84 Å, (D) cluster from family 1 (centroid) – RMSD 3.93 Å, (E) cluster from family 2 (lowest energy) – RMSD 4.86 Å, (F), cluster from family 2 (centroid) – RMSD 4.84 Å.

Article Snippet: Subsequently, 2-fold serial dilutions of PD-1-Fc (Sino Biological Company, China, #10084-H08H-B) in PBS-T, at concentrations from 8.00 to 0.5 µg/ml, were added to the precoated wells.

Techniques: