초록 ▼

An assay method and compositions are provided for determining the presence of an analyte in a sample. The analyte is a member of an immunological pair (mip) of immunogens－ligand and receptor. The method has two basic elements: a solid surface to which one of the members of the immunological pair is

An assay method and compositions are provided for determining the presence of an analyte in a sample. The analyte is a member of an immunological pair (mip) of immunogens－ligand and receptor. The method has two basic elements: a solid surface to which one of the members of the immunological pair is bonded and a signal producing system, which includes a catalytic member bonded to a mip, which signal producing system results in a measurable signal on said solid surface related to the amount of analyte in the medium. The signal generating compound is produced without separation of the catalyst labeled mip bound to the solid surface from the catalyst labeled mip free in solution. In a preferred embodiment, an enzyme is bonded to a mip which acts in conjunction with a solute to produce a signal generating product which binds preferentially to the solid surface when the enzyme is bound to the surface, resulting in a signal which is readily differentiated from signal generating compound produced by the catalyst and solute in the bulk solution.

대표청구항 ▼

A method for detecting the presence of an analyte in a sample suspected of containing said analyte, where said analyte is a member of an immunological pair (mip) consisting of ligand and homologous antiligand; said method involving (1) the partitioning of an enzyme bound to a mip－enzyme-bound-mip－be

A method for detecting the presence of an analyte in a sample suspected of containing said analyte, where said analyte is a member of an immunological pair (mip) consisting of ligand and homologous antiligand; said method involving (1) the partitioning of an enzyme bound to a mip－enzyme-bound-mip－between a surface and a liquid phase, where said partitioning is through the intermediacy of ligand-antiligand binding to a mip-bound-surface in relation to the amount of analyte in said sample;, said surface being porous and permitting approach and binding of enzyme-bound-mip to mip of said mip-bound-surface, either directly or in combination with said ligand and (2) the change in intensity of a detectible signal from a surface resulting from the change in concentration of a signal generating compound associated with said surface, said change in intensity being related to the amount of reaction product produced by said enzyme bound to said surface; said method comprising: (a) combining in an aqueous assay medium; (1) said sample; (2) mip-bound-surface, substantially immersed in said medium wherein substantially all of said mip-bound-surface is uniformly contacted with said sample in said medium; (3) enzyme-bound-mip; and (4) the remaining members of the signal producing system, which system includes at least a second enzyme other than said enzyme-bound-mip, wherein the product of one enzyme is the substrate of the other enzyme, and a solute which is capable of undergoing a catalyzed reaction to produce a product which results in a change in amount of a signal generating compound associated with said surface and capable of producing a detectible signal, (b) waiting a sufficient time for enzyme-bound-mip to diffuse to said surface and at least a portion of said enzyme-bound-mip to bind to said surface through the intermediacy of ligand-antiligand binding and for a change in the amount of signal generating compound associated with said surface in relation to the amount of analyte in said sample; (c) determining the intensity of said detectible signal at said surface as a function of the amount of analyte in said sample.