G - Physics – 01 – N
Patent
G - Physics
01
N
167/44
G01N 33/545 (2006.01) G01N 33/531 (2006.01) G01N 33/537 (2006.01) G01N 33/569 (2006.01)
Patent
CA 1279009
Abstract of the Disclosure An immunoassay on which a thermally induced phase separation is used to effect the separation of specifically bound reactants from free reactants is disclosed. A first reactant is conjugated to a temperature-sensitive polymer to form a polymer/reactant conjugate, and a second reactant is conjugated to a reporter to form a reporter/reactant conjugate. The polymer/reactant, reporter/reactant, and biological fluid samples suspected of containing the analyte are admixed in solu- tion at a temperature other than that at which the polymer will precipitate. Specific binding is allowed to occur, thereby forming ternary complex. The salt concentration of the adjusted solution is then adjusted to a concentration suffi- cient to cause the complex to precipitate from the solution, the amount of reporter activity in the precipitated complex or in the solution measured and the present and/or concentration of the analyte therefrom determined. Alternatively, the first reactant may be conjugated to a monomer and subsequently copoly- merized with additional monomers to yield a temperature- sensitive copolymer. As a further alternative, an activated, thermally reversible copolymer characterized by a lower critical solution temperature is produced by copolymerization of a monomer which produces a polymer having a lower critical solution temperature, and a second monomer. The second monomer is either a vinyl substituted monomer capable of binding to a specific binding partner, or a monomer which must be activated after polymerization to produce a functionality capable of binding to a specific binding partner. Multiple analyses may also be performed on a single sample by choosing a variety of polymers, each polymer having a different specific binding partner conjugated thereto and a different critical solution temperature. By altering the temperature and/or the salt oncentration of the solution incrementally, the reporter associated with each of the complexes precipitated with each temperature or concentration increment may be measured, and the presence and/or concentration of each of the analytes determined.
508267
Cole Carolann
Hoffman Allan S.
Houghton Raymond L.
Monji Nobuo
Priest John H.
Genetic Systems Corporation
Mccarthy Tetrault Llp
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