Biosensor device and method
First Claim
1. A biosensor apparatus for detecting a binding event between a ligand and ligand-binding agent, comprisingmeans defining a biosensor surface, carried on said biosensor surface, two-subunit heterodimer complexes composed of first and second peptides that together form an α
- -helical coiled-coil heterodimer, where said first peptide is attached to the biosensor surface, a ligand covalently attached to the second peptide in the two-subunit heterodimer complexes, accessible for binding by a ligand-binding agent, means for introducing onto the biosensor surface, an analyte selected from the group consisting of a ligand-binding agent, and a ligand or ligand analog capable of competing with a ligand-binding agent for binding to said ligand, said apparatus also including a ligand-binding agent when the analyte is a ligand or ligand analog, and means for detecting the binding of a ligand-binding agent to the ligand on the biosensor surface.
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Abstract
A biosensor apparatus for detecting a binding event between a ligand and receptor. The apparatus includes a biosensor surface and surface-bound two-subunit heterodimer complexes composed of first and second, preferably oppositely charged peptides that together form an α-helical coiled-coil heterodimer. The first peptide is attached to the biosensor surface, and the second peptide carries the ligand, accessible for binding by a ligand-binding agent. Binding of anti-ligand binding agent to the surface-bound ligand is detected by a suitable detector. A ligand-specific biosensor surface can be readily prepared from a universal template containing the first charged peptide, by addition of a selected ligand attached to the second peptide.
73 Citations
8 Claims
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1. A biosensor apparatus for detecting a binding event between a ligand and ligand-binding agent, comprising
means defining a biosensor surface, carried on said biosensor surface, two-subunit heterodimer complexes composed of first and second peptides that together form an α - -helical coiled-coil heterodimer, where said first peptide is attached to the biosensor surface,
a ligand covalently attached to the second peptide in the two-subunit heterodimer complexes, accessible for binding by a ligand-binding agent, means for introducing onto the biosensor surface, an analyte selected from the group consisting of a ligand-binding agent, and a ligand or ligand analog capable of competing with a ligand-binding agent for binding to said ligand, said apparatus also including a ligand-binding agent when the analyte is a ligand or ligand analog, and means for detecting the binding of a ligand-binding agent to the ligand on the biosensor surface.
- -helical coiled-coil heterodimer, where said first peptide is attached to the biosensor surface,
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2. An array device for detecting each of a plurality of binding events between a ligand and ligand-binding agent, comprising
a substrate defining an array of surface regions, carried on each of said regions, two-subunit heterodimer complexes composed of first and second peptides that together form an α - -helical coiled-coil heterodimer, where said first peptide is attached to the biosensor surface, and
a ligand covalently attached to the second peptide in the two-subunit heterodimer complexes, accessible for binding by a ligand-binding agent. - View Dependent Claims (3, 4, 5, 6, 7, 8)
said detector includes means for irradiating said monolayer with a light beam, and means for detecting a change in the optical characteristics of the monolayer produced by binding of ligand-binding agent to said ligand.
- -helical coiled-coil heterodimer, where said first peptide is attached to the biosensor surface, and
Specification