Patent ID: 7255998
Filing Date: 2007-08-14
Classification: G01N,Y10S

Abstract:
1. A time-resolved fluoroimmunoassay (TR-FIA) method for detecting a cytokine in a biological fluid sample, comprising: subjecting the biological fluid sample to a heat treatment under non-denaturing temperature conditions of about 55° C. for the cytokine; forming a composite by (a) binding a first antibody, including a portion bound to a solid phase and a region bindable to a cytokine, to a solid phase; (b) adding the sample containing the cytokine; (c) binding a second antibody, including a region bindable to the cytokine and a portion to which biotin is bound, to the cytokine; (d) adding a conjugate including streptoavidin or avidin and a fluorescent structural portion capable of being complexed with a lanthanoid metal ion; and (e) adding a lanthanoid metal ion to bind the conjugate of (d), the composite being formed on the solid phase; and measuring fluorescence of the fluorescent structural portion which has been complexed with the lanthanoid metal ion, wherein the method comprises a step of washing after each of steps (a) to (c); and wherein the cytokine is a cytokine belonging to the chemokine family, and wherein the fluorescent structural portion is represented by General Formula (I): (where R is a residue which is a functional group capable of forming a covalent bond with a protein; Ar is a hydrocarbon group having a conjugated double bond system; n is an integer equal to or greater than 1; and X is a fluorine atom or a group represented by General Formula (II):