A method for detecting a cytokine in a biological fluid sample with a high
sensitivity is provided. A time-resolved fluoroimmunoassay (TR-FIA)
method including a step of forming on a solid phase a composite in which
a cytokine is captured and which includes a fluorescent structural
portion which has been complexed with a lanthanoid metal ion, and
measuring fluorescence of the fluorescent structural portion. The
composite is formed of a structure in which (a) a first antibody
including a portion bound to a solid phase and a region bindable to a
cytokine; (b) the cytokine; (c) a second antibody including a region
bindable to the cytokine and a portion to which biotin is bound; (d) a
conjugate including streptoavidin or avidin and a fluorescent structural
portion capable of being complexed with a lanthanoid metal ion; and (e)
the lanthanoid metal ion are bound. The fluorescent structural portion is
represented by General Formula (I):
R--Ar--C(.dbd.O)--CH.sub.2--C(.dbd.O)--C.sub.nF.sub.2n--X.