Calibration device for heterogeneous immunoassay
AssigneeSyntex (U.S.A.) Inc.
InventorsN. Chung Sizto, Cynthia G. Roux
Life of this patent
Estimating the term…
Abstract
A device is disclosed for use in a simultaneous calibration assay method wherein a conjugate of a catalyst and a member of a specific binding pair ("mip") is employed. The present device comprises a surface to which a receptor is non-diffusively bound. The receptor is capable of specific binding to the conjugate of the catalyst and mip and is substantially incapable of binding to the catalyst or the mip apart from the conjugate. The surface may be an integral part of, or separate from, a support.
Claims19
In a method for determining the presence in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"), said method comprising the step of:
- (a) contacting with said sample (i) at least one enzyme including an enzyme bound to a mip ("enzyme-bound-mip") and (ii) a solute which is catalytically transformed by an enzyme bound to a mip-containing measurement first surface to produce a change in a detectable signal at said first surface in proportion to the amount of enzyme-bound-mip bound to said first surface, wherein said enzyme-bound-mip binds to said first surface in proportion to the amount of analyte in said sample, and (iii) a calibration second surface, adjacent to said first surface to which second surface enzyme becomes bound in an amount which provides substantially predetermined ratios to the amount of said enzyme bound to said first surface, whereby the intensity of the signal at said second surface compared to the intensity of signal at said first surface is related to the amount of analyte in said sample, and
- (b) determining said signal,
- the improvement which comprises employing on said second surface a receptor for said enzyme-bound-mip, said receptor being capable of specific binding to said enzyme-bound-mip and being substantially incapable of binding to said enzyme or said mip part from said enzyme-bound-mip.
In a method for determining the presence in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"), said method comprising the steps of
- (a) contacting with said sample (i) a signal producing system having at least two enzymes, including one enzyme bound to a mip ("enzyme-bound-mip"), and (ii) a solute dye precursor which is catalytically transformed to an insoluble dye by one of said enzymes bound to a mip-containing measurement first surface, said insoluble dye producing a change in a detactable signal at said first surface in proportion to the amount of analyte, where said mip at said first surface provides for binding of enzyme-bound-mip through mip complex formation to said first surface and said enzyme-bound-mip binds to said first surface in proportion to the amount of analyte in said sample, and (iii) a calibration second surface, adjacent to said first surface, to which second surface enzyme of said enzyme-bound-mip becomes bound through mip complex formation in an amount which provides substantially predetermined ratios to the amount of enzyme-bound-mip on said first surface, where the intensity of the signal at said second surface compared to the intensity of signal at said first surface is related to the amount of analyte in said sample, and
- (b) determining said signal,
- the improvement which comprises employing on said second surface a receptor for said enzyme-bound-mip, said receptor being capable of specific binding to said enzyme-bound-mip and being substantially incapable of binding to said enzyme or said mip apart from said enzyme-bound-mip.
A method for determining the presence in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"), which method comprises the steps of
- (a) contacting an enzyme bound to a mip ("enzyme-bound-mip"), a solute which is catalytically transformed by an enzyme, and said sample with a measurement surface containing a mip and an enzyme different from that of the enzyme-bound-mip, and a calibration surface, adjacent to said measurement surface, containing enzyme different from that of the enzyme-bound-mip and an antibody specific for said enzyme-bound-mip, said antibody being capable of specific binding to said enzyme-bound-mip and being substantially incapable of binding to said enzyme or said mip apart from said enzyme-bound-mip, wherein the enzymes are related by the product of one being the substrate of the other, and
- (b) measuring a change in a detactable signal at said measurement surface, wherein said enzyme-bound-mip becomes bound to said calibration surface in an amount which provides substantially predetermined ratios to the amount of said enzyme bound to said measurement surface, whereby the intensity of the signal at said calibration surface compared to the intensity of signal at said measurement surface is related to the amount of analyte in said sample.
A internally calibrated diagnostic device comprising a support, a measurement surface of a porous material, a calibration surface of a porous material in close proximity to said measurement surface, a member of a specific binding pair ("mip") non-diffusively bound to said measurement surface, and a receptor non-diffusively bound to said calibration surface, said receptor being capable of specific binding to a conjugate of an enzyme and a mip but substantially incapable of binding to said enzyme or said mip apart from said conjugate.
Dependencies are inferred from the claim text.