Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Schematic representation of the principle of SARS-CoV-2-RBD neutralizing ELISA assay2.2. Assay Optimization

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Standard curve of anti-SARS-COV-2-RBD nAbs for the in-house developed SARS-COV-2-RBD neutralizing ELISA assay

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Mean ± standard deviation of OD values of different anti-SARS-COV-2-RBD neutralizing antibody standards generated by the designed neutralizing ELISA kit

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Standard Deviation, Generated, Enzyme-linked Immunosorbent Assay, Concentration Assay, Negative Control, Positive Control

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Cut-off value results of the in-house neutralizing ELISA; A) Optimized ROC analysis cut-off value of positive and B) Concentration and log concentration limits for the optimal established cut-off in this assay

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Receiver operating characteristic curve analysis for the optimal cut-off value for detecting the positive anti-SARS-COV-2-RBD neutralizing antibodies using the in-house developed neutralizing ELISA kit

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Serum panels using the in-house designed SARS-COV-2-RBD neutralizing ELISA assay; A) Positive control samples for the assay validation and B) Negative control samples for the assay validation

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Enzyme-linked Immunosorbent Assay, Positive Control, Biomarker Discovery, Negative Control, Concentration Assay

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Agreement of the results between the in-house SARS-COV-2-RBD neutralizing ELISA assay and gold standard test at the in-house established cut-off

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Archives of Razi Institute

Article Title: Application and Validation of SARS-CoV-2 RBD Neutralizing ELISA Assay

doi: 10.22092/ARI.2021.356677.1890

Figure Lengend Snippet: Validation parameters of the in-house SARS-COV-2-RBD neutralizing ELISA assay

Article Snippet: To immobilize the hACE2 protein at 400 ng/well in 100 μl of 1X working Coating Buffer on the microtiter plates wells, a calculated volume from Stock solution vial of Recombinant His-Tag-Human ACE2 protein in a concentration of 2.8 mg/ml (Raybiotech) was diluted immediately before usage with the prepared 1X working coating buffer from 5X stock ELISA Plate Coating Buffer (Elabscience) to a required volume at the appropriate concentration (4 ng/μl), which was determined in the initial experiments.

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Experimental Medicine

Article Title: VISTA is an activating receptor in human monocytes

doi: 10.1084/jem.20201601

Figure Lengend Snippet: Binding and specificity of novel anti-VISTA mAbs. (A) VISTA mAb1 and VISTA mAb2 binding to plate-bound recombinant human (hu) PD-L1, huB7H5, or murine (mu) B7H5 by ELISA. (B) Binding kinetics of mAb1 and mAb2 to human and murine VISTA by SPR. No binding (N.B.) was observed for mAb2 to murine VISTA. (C) Representative Biacore sensorgram plots from simultaneous binding assay with mAb1 or mAb2 captured on the surface. Antigen binding to surface mAb is shown, followed by injection of solution mAb (soln). RU, reference units. (D) Summarized FACS staining results that quantify the percentage of VISTA + cells in human whole blood. n = 6 independent donors. T N , naive T cells; T Ag-Ex , antigen-experienced T cells. (E) FACS-based median fluorescence intensity (MFI) of isotype control, mAb1, or mAb2 binding to CD14 + PBMCs while in the presence of titrated recombinant His-tagged VISTA (VISTA-His; green triangles or blue circles) or recombinant His-tagged PD-L1 (PD-L1 His; purple triangles or red squares). A–E are representative of three independent experiments.

Article Snippet: The ELISA Substrate Reagent Pack (DY999), ELISA plate-coating buffer (DY006), reagent diluent concentrate 2 (DY995), and Stop Solution (DY994) were purchased from R&D Systems.

Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Injection, Staining, Fluorescence, Control

Journal: The Journal of Experimental Medicine

Article Title: VISTA is an activating receptor in human monocytes

doi: 10.1084/jem.20201601

Figure Lengend Snippet: Characterization of VISTA.COMP and PD-L1.COMP binding on human PBMCs. Related to . (A) Left unstained (gray) or stained with PD-L1.COMP (blue) or VISTA.COMP (red) protein and markers that identify immune cell subtypes, including T cells (CD3 + ), NK cells (CD56 + ), NK T cells (CD3 + CD56 + ), and monocytes (CD14 + SSC hi ) in the presence of 0.5% paraformaldehyde. FSC, forward scatter; SSC, side scatter. (B) Gating strategy for PBMC immune cell identification and T cell subtyping. (C) PD-L1.COMP and VISTA.COMP (negative control) binding to plate-bound, recombinant human (hu) PD1-Fc by ELISA. A and C were performed in two independent experiments.

Article Snippet: The ELISA Substrate Reagent Pack (DY999), ELISA plate-coating buffer (DY006), reagent diluent concentrate 2 (DY995), and Stop Solution (DY994) were purchased from R&D Systems.

Techniques: Binding Assay, Staining, Negative Control, Recombinant, Enzyme-linked Immunosorbent Assay