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Lifetime-resolved assay procedures

  • US 4,822,733 A
  • Filed: 05/28/1985
  • Issued: 04/18/1989
  • Est. Priority Date: 05/28/1985
  • Status: Expired due to Term
First Claim
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1. In a luminescent assay for detection of an analyte in a sample wherein a reaction mixture is formed containing the analyte and a photophore-labelled probe capable of a specific association with the analyte and wherein the presence of the analyte is determined by luminescent emission of a photophore upon excitation of the reaction mixture by a modulated light source, the improvement comprising:

  • (1) employing in said reaction mixture a first photophore-labelled probe capable of a specific association with said analyte, the photophore of which has a first emissive lifetime, anda second photophore-labelled probe capable of a specific association with said analyte, the photophore of which has a second emissive lifetime,one of the photophores having a significantly longer emissive lifetime than the other photophore,said one photophore having a significantly longer emissive lifetime being excitable by said modulated light source to an excited state from which energy may be transferred to said other photophore having a significantly shorter emissive lifetime when in sufficient proximity thereto, thereby causing excitation and emisssion of said other photophore,the association of said analyte with said first probe and said second probe resulting in said photophores of said first and second probes being in sufficient proximity to each other to allow the excitation of said one photophore having a significantly longer emissive lifetime by said modulated light source to result in transfer of energy to said other photophore having a significantly shorter emissive lifetime causing excitation of and emission by said other photophore;

    (2) exciting the reaction mixture with said modulated light source;

    (3) monitoring the reaction mixture to determine the luminescent emission of said other photophore having a significantly shorter emissive lifetime as excited by energy transfer from said one photophore excited by said modulated energy source, at a time beyond the lifetime of emission of said other photophore having a significantly shorter emissive lifetime relative to the time of each excitation of said reaction mixture by said modulated light source and(4) using the determination of luminescent emission of said other photophore obtained by said monitoring step to determine the amount of analyte present in said sample.

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