Method for analysis for a member of an immunological pair using a test surface; kit and test surface material therefor

Claim 1

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 a catalyst bound to a mlp--catalyst-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; and (2) the change in concentration of a signal generating compound associated with said surface, said change in concentration affecting the amount of reaction product produced by said catalyst bound to said surface; by a) combining in an aqueous assay medium, 1) said sample; 2) mip-bound-surface; 3) catalyst-bound-mip; and 4) the remaining members of the signal producing system, which system includes at least one catalyst including said catalyst-bound-mip, and a solute which is capable of undergoing a catalyzed reaction to produce a product which results in the formation or destruction of a signal generating compound associated with said surface and capable of producing a detectible signal, with the proviso that said signal producing system is completed not later than about the time of addition of said catalyst-bound-mip to said surface, when said signal producing system consists essentially of said catalyst-bound-mip and its substrates; b) waiting a sufficient time for at least a portion of said catalyst-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; and c) determining the amount of detectible signal at said surface as a function of the presence of analyte in said sample.

Claim 2

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 a catalyst bound to a mlp--catalyst-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; and (2) the change in concentration of a signal generating compound associated with said surface, said change in concentration affecting the amount of reaction product produced by said catalyst bound to said surface; by a) combining in an aqueous assay medium, 1) said sample; 2) mip-bound-surface; 3) catalyst-bound-mip; and 4) the remaining members of the signal producing system, which system includes at least one catalyst including said catalyst-bound-mip, and a solute which is capable of undergoing a catalyzed reaction to produce a product which results in the formation or destruction of a signal generating compound associated with said surface and capable of producing a detectible signal, with the proviso that said signal producing system is completed not later than about the time of addition of said catalyst-bound-mip to said surface, when said signal producing system consists essentially of said catalyst-bound-mip and its substrates; b) waiting a sufficient time for at least a portion of said catalyst-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; and c) determining the amount of detectible signal at said surface as a function of the presence of analyte in said sample.

Claim 6

A method for detecting the presence of a ligand analyte in a sample suspected of containing said ligand where said ligand 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; and (2) the change in concentration of a signal generating compound associated with said surface, where said signal generating compound is an insoluble dye, said change in concentration being in relation to the amount of insoluble dye 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, wherein said mip-bound-surface is substantially uniformly contacted with said surface; b) incubating for sufficient time for ligand analyte to bind to said mip-bound-surface; c) after sufficient time for said incubation, adding substantially concurrently enzyme-bound-mip; and d) remaining members of said signal producing system, which system includes at least said enzyme of said enzyme-bound-mip and a solute which is capable of undergoing a catalyzed reaction to produce a product which results in the formation of a signal generating compound, which is an insoluble dye capable of producing a detectible signal, e) incubating for sufficient time for said insoluble dye to precipitate on said surface in proportion to the amount of analyte in said sample; and f) determining the amount of said detectible signal on said surface as a function of the amount of analyte in said sample.

Claim 7

A method for detecting the presence of a ligand analyte in a sample suspected of containing said ligand where said ligand 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; and (2) the change in concentration of a signal generating compound associated with said surface, where said signal generating compound is an insoluble dye, said change in concentration being in relation to the amount of insoluble dye 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, wherein said mip-bound-surface is substantially uniformly contacted with said surface; b) incubating for sufficient time for ligand analyte to bind to said mip-bound-surface; c) after sufficient time for said incubation, adding substantially concurrently enzyme-bound-mip; and d) remaining members of said signal producing system, which system includes at least said enzyme of said enzyme-bound-mip and a solute which is capable of undergoing a catalyzed reaction to produce a product which results in the formation of a signal generating compound, which is an insoluble dye capable of producing a detectible signal, e) incubating for sufficient time for said insoluble dye to precipitate on said surface in proportion to the amount of analyte in said sample; and f) determining the amount of said detectible signal on said surface as a function of the amount of analyte in said sample.

Claim 9

A method according to any one of the preceding claims wherein said signal producing system includes a scavenger in the liquid medium which reacts with a component of said signal producing system to inhibit operation of said signal producing system in said liquid phase.