Method and reagents for binding chemical analytes to a substrate surface, and related analytical devices and diagnostic techniques
AssigneeHewlett-Packard Company
InventorsMay Tom-Moy, Joel Myerson, Karla M. Robotti
Life of this patent
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Abstract
A method is provided for detecting an analyte of interest in a sample. The method involves binding the analyte to the surface of a substrate through a biotin-biotin binding protein interaction, contacting the surface-bound analyte with a quantitatively detectable analyte-binding moiety that binds thereto, measuring the quantity of detectable moiety bound to the substrate surface and deriving therefrom the quantity of analyte in solution. A preferred use for the present method is in conjunction with a piezoelectric surface transverse wave device. Novel reagents useful for carrying out the inventive method are provided as well.
Claims23
A method for binding an environmental analyte having a molecular weight of less than about 1000 to a surface of a piezoelectric substrate, comprising:
- coating the surface with a layer of a specific binding material comprised of a biotin-binding protein to provide a reactive surface;
- covalently binding the analyte, either directly or indirectly, to biotin, to provide a biotin-analyte complex;
- and
- coating the reactive surface with the biotin-analyte complex such that the analyte is bound thereto.
A method for quantitating an environmental analyte having a molecular weight of less than about 1000, comprising:
- (a) providing a piezoelectric substrate comprising surface-bound analyte by
- (i) coating the surface with a layer of a protein which specifically binds biotin, to provide a reactive surface,
- (ii) covalently binding the analyte, either directly or indirectly, to biotin, to provide a biotin-analyte complex, and
- (iii) coating the reactive surface with the biotin-analyte complex to provide the surface with bound analyte;
- (b) contacting the surface-bound analyte with
- (i) a specific binding partner which specifically binds to the analyte and an analog thereof, and
- (ii) a first liquid sample suspected of containing the analyte,
- such that the formation of a first specific binding complex that comprises the specific binding partner and the surface bound analyte produces a first detectable mass on the piezoelectric substrate, wherein the first detectable mass is inversely proportional to the amount of the analyte in the first liquid sample;
- (c) contacting the surface-bound analyte with
- (i) the specific binding partner, and
- (ii) a second liquid sample containing a standard comprising a known concentration of the analyte or the analog thereof, such that the formation of a second specific binding complex that comprises the specific binding partner and the surface bound analyte produces a second detectable mass on the piezoelectric substrate, wherein the second detectable mass is inversely proportional to the amount of the standard in the second liquid sample;
- and
- (d) determining the quantity of the analyte in the first liquid sample by correlating the first detectable mass produced on the piezoelectric substrate by the first specific binding complex to the second detectable mass produced on the piezoelectric substrate by the second specific binding complex.
A biotin-analyte complex having the structural formula ##STR10## wherein X is a C.sub.1 -C.sub.12 alkylene linking group substituted with 0 to 4 substituents selected from the group consisting of lower alkyl, hydroxyl, halogen and amino, optionally containing 1 to 4 --O--, --NH--, --CONH-- or --(CO)-- linkages and A is an analyte having a molecular weight of less than about 1000.
Dependencies are inferred from the claim text.