Assay to detect HCV receptor binding
AssigneeNovartis Vaccines and Diagnostics, Inc.
InventorSergio Abrignani
Life of this patent
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Abstract
Identification of HCV receptor target cells using HCV receptor-binding ligands and cell separation by flow cytofluorimetry is described. HCV receptor target cells are employed to conduct assays for HCV receptor-binding ligands in order to identify potential HCV vaccine candidates. HCV receptor target cells are used to measure antibody neutralisation to monitor vaccine development, as a diagnostic of HCV infection and to develop neutralising antibodies for passive immunisation.
Claims12
A method for determining the ability of an HCV receptor binding ligand to interfere with the ability of HCV E2 to bind to an HCV receptor target cell, said method comprising:
- contacting MOLT-4 cells with (i) an HCV E2, and (ii) a putative HCV receptor binding ligand, under conditions that allow binding of the HCV E2 and/or the putative receptor binding ligand to the MOLT-4 cells to form HCV E2 MOLT-4 cell complexes and/or HCV receptor binding ligand MOLT-4 cell complexes;
- removing unbound HCV E2 and HCV receptor binding ligands;
- adding a detectable anti-E2 antibody to label those MOLT-4 cells which have bound HCV E2;
- detecting the amount of labeled MOLT-4 cells as a measurement of the ability of an HCV receptor binding ligand to prevent binding of HCV E2 with an HCV receptor target cell.
A method for determining the ability of an HCV receptor binding ligand to interfere with the ability of HCV E2 to bind to an HCV receptor target cell, said method comprising:
- contacting MOLT-4 cells with (i) HCV E2384-715, wherein the HCV E2 is produced recombinantly in a mammalian cell, and (ii) a putative HCV receptor binding ligand, under conditions that allow binding of the HCV E2 and/or the putative receptor binding ligand to the MOLT-4 cells to form HCV E2 MOLT-4 cell complexes and/or HCV receptor binding ligand MOLT-4 cell complexes;
- removing unbound HCV E2384-715 and HCV receptor binding ligands;
- adding a detectable anti-E2 antibody to label those MOLT-4 cells which have bound HCV E2384-715;
- detecting the amount of labeled MOLT-4 cells as a measurement of the ability of an HCV receptor binding ligand to prevent binding of HCV E2384-715 with an HCV receptor target cell using flow cytometry.
A method for determining the ability of an HCV receptor binding ligand to interfere with the ability of an HCV E1/E2 to bind to an HCV receptor target cell, said method comprising:
- contacting MOLT-4 cells with (i) an HCV E1/E2, and (ii) a putative HCV receptor binding ligand, under conditions that allow binding of the HCV E1/E2 and/or the putative receptor binding ligand to the MOLT-4 cells to form HCV E1/E2 MOLT-4 cell complexes and/or HCV receptor binding ligand MOLT-4 cell complexes;
- removing unbound HCV E1/E2 and HCV receptor binding ligands;
- adding a detectable anti-E2 antibody to label those MOLT-4 cells which have bound HCV E1/E2;
- detecting the amount of labeled MOLT-4 cells as a measurement of the ability of an HCV receptor binding ligand to prevent binding of HCV E1/E2 with an HCV receptor target cell.
A method for determining the ability of an HCV receptor binding ligand to interfere with the ability of HCV E1/E2 to bind to an HCV receptor target cell, said method comprising:
- contacting MOLT-4 cells with (i) HCV E1/E2199-745, wherein the HCV E1/E2199-745 is produced recombinantly in a mammalian cell, and (ii) a putative HCV receptor binding ligand, under conditions that allow binding of the HCV E1/E2199-745 and/or the putative receptor binding ligand to the MOLT-4 cells to form HCV E1/E2199-745 MOLT-4 cell complexes and/or HCV receptor binding ligand MOLT-4 cell complexes;
- removing unbound HCV E2384-715 and HCV receptor binding ligands;
- adding a detectable anti-E2 antibody to label those MOLT-4 cells which have bound HCV E2384-715;
- detecting the amount of labeled MOLT-4 cells as a measurement of the ability of an HCV receptor binding ligand to prevent binding of HCV E2384-715 with an HCV receptor target cell using flow cytometry.
Dependencies are inferred from the claim text.