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US 8,617,808 B2Granted
Cultispot assay
AssigneeMabtech AB
InventorsSten Braesch-Andersen, Staffan Paulie
Life of this patent
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Abstract
The invention provides a method for detecting the production of one or more analytes by cells in a sample. The method uses one or more immobilized binding proteins to capture one or more analytes released from cells. The, or each, binding protein is localized at one or more discrete locations on a solid surface within a well. Cells are introduced into the well and cultured under conditions suitable for release of the analyte(s) and for binding of the analyte(s) to the binding protein(s). Restricting the binding protein(s) to one or more discrete locations within the well, rather than coating the entire surface of the well, results in increased sensitivity of the assay.
Claims18
A method for detecting the production of more than one analyte by cells in a sample, which method comprises:
- (a) introducing a sample of cells into a well, wherein
- (i) a first binding protein which specifically binds to a first analyte;
- (ii) a second binding protein which specifically binds to a second analyte;
- and
- (iii) optionally one or more further binding protein which specifically binds to one or more further analyte;
- are each immobilized at one or more discrete locations on a solid surface within the well, wherein said discrete locations are spots;
- (b) culturing said sample of cells under conditions suitable for:
- (i) release of said first analyte, said second analyte and optionally said one or more further analyte by said cells;
- and
- (ii) binding of said first analyte to said first binding protein, binding of said second analyte to said second binding protein and optionally binding of said one or more further analyte to said one or more further binding proteins;
- and
- (c) determining whether or not said first analyte is bound to said first binding protein, whether or not said second analyte is bound to said second binding protein, and whether or not said one or more further analyte is bound to said one or more further binding proteins
- thereby detecting production by a sample of cells of more than one analyte.
10 more elements
A method for detecting the production of more than one analyte by cells in a sample, which method comprises:
- (a) introducing a sample of cells into a well, wherein
- (i) a first binding protein which specifically binds to a first analyte;
- (ii) a second binding protein which specifically binds to a second analyte;
- and
- (iii) optionally one or more further binding protein which specifically binds to one or more further analyte;
- are each immobilized at one or more discrete locations on a solid surface within the well;
- (b) culturing said sample of cells under conditions suitable for:
- (i) release of said first analyte, said second analyte and optionally said one or more further analyte by said cells;
- and
- (ii) binding of said first analyte to said first binding protein, binding of said second analyte to said second binding protein and optionally binding of said one or more further analyte to said one or more further binding proteins,
- wherein the cells are cultured in the presence of a first agent capable of inducing or inhibiting production of said first analyte, said second analyte and/or said one or more further analyte and a second agent which enhances detection of one or more of said first analyte, said second analyte or said one or more further analyte, wherein said first agent and/or said second agent is immobilized in said well,
- (c) determining whether or not said first analyte is bound to said first binding protein, whether or not said second analyte is bound to said second binding protein, and whether or not said one or more further analyte is bound to said one or more further binding proteins,
- thereby detecting production by a sample of cells of more than one analyte.
10 more elements
A method for detecting the production of one or more analytes by cells in a sample, which method comprises:
- (a) introducing a sample of cells into a well, wherein
- (i) a first binding protein which specifically binds to a first analyte;
- and
- (ii) optionally a second binding protein which specifically binds to a second analyte;
- are each immobilized in separate spots arrayed on a solid surface within the well;
- (b) culturing said sample of cells under conditions suitable for:
- (i) release of said first analyte and optionally said second analyte by said cells;
- and
- (ii) binding of said first analyte to said first binding protein and optionally binding of said second analyte to said second binding protein;
- and
- (c) determining whether or not said first analyte is bound to said first binding protein and whether or not said second analyte is bound to said second binding protein,
- thereby detecting production by a sample of cells of one or more analyte.
9 more elements
A method for detecting the production of more than one analyte by cells in a sample, which method comprises:
- (a) introducing a sample of cells into a well, wherein
- (i) a first binding protein which specifically binds to a first analyte;
- (ii) a second binding protein which specifically binds to a second analyte;
- and
- (iii) optionally one or more further binding protein which specifically binds to one or more further analyte;
- are each immobilized at one or more discrete locations on a solid surface within the well, where the discrete locations are spots;
- (b) culturing said sample of cells under conditions suitable for:
- (i) release of said first analyte, said second analyte and optionally said one or more further analyte by said cells;
- and
- (ii) binding of said first analyte to said first binding protein, binding of said second analyte to said second binding protein and optionally binding of said one or more further analyte to said one or more further binding protein;
- and
- (c) determining whether or not said first analyte is bound to said first binding protein, whether or not said second analyte is bound to said second binding protein, and whether or not said one or more further analyte is bound to said one or more further binding protein,
- thereby detecting production by a sample of cells of more than one analyte.
10 more elements
A method for detecting the production of more than one analyte by cells in a sample, which method comprises:
- (a) introducing a sample of cells into a well, wherein
- (i) a first binding protein which specifically binds to a first analyte;
- (ii) a second binding protein which specifically binds to a second analyte;
- and
- (iii) optionally one or more further binding protein which specifically binds to one or more further analyte;
- are each immobilized at one or more discrete locations on a solid surface within the well;
- (b) culturing said sample of cells under conditions suitable for:
- (i) release of said first analyte, said second analyte and optionally said one or more further analyte by said cells;
- and
- (ii) binding of said first analyte to said first binding protein, binding of said second analyte to said second binding protein and optionally binding of said one or more further analyte to said one or more further binding protein;
- wherein said cells are cultured in the presence of a first agent capable of inducing or inhibiting production of said first analyte, said second analyte and/or said one or more further analyte and a second agent which enhances detection of one or more of said first analyte, said second analyte or said one or more further analyte, wherein said first agent and/or said second agent are immobilized at one or more discrete locations in said well, and
- (c) determining whether or not said first analyte is bound to said first binding protein, whether or not said second analyte is bound to said second binding protein, and whether or not said one or more further analyte is bound to said one or more further binding protein,
- thereby detecting production by a sample of cells of more than one analyte.
10 more elements
Dependencies are inferred from the claim text.