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Immunological detection using two detectable labels

  • US 5,723,304 A
  • Filed: 02/27/1995
  • Issued: 03/03/1998
  • Est. Priority Date: 08/03/1992
  • Status: Expired due to Fees
First Claim
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1. In a competitive assay method for immunological detection in a sample of the presence or concentration of an analyte species, wherein (a) a primary species comprises an analyte analog;

  • (b) the primary species provides or is associated with a first detectable species;

    (c) a primary species reaction partner is provided which is capable of undergoing a specific binding reaction in a primary immune reaction mixture with the analyte species, if present, or with the analyte analog;

    (d) both the sample and the primary species are exposed to and incubated with the primary species reaction partner;

    (e) means are provided for separating the primary species from the reaction mixture; and

    (f) a signal from the first detectable species is detected by a signal-detecting means, the improvement comprising;

    (i) causing a second detectable specics to become associated with a support material via a secondary species and a secondary species reaction partner, said secondary species (A) being a species which does not itself undergo a specific binding reaction with the analyte species or with the analyte analog, (B) providing or being associated with the second detectable species, (C) undergoing a specific binding reaction with the secondary species reaction partner which is provided on or by, or associated with, the support material, and (D) being or becoming linked to the primary species reaction partner; and

    (ii) if the analyte analog becomes bound to the primary species reaction partner, causing the first detectable species to become associated with the support material via linkage of the primary species reaction partner to the secondary species, with signals from said first and second detectable species being detectable by the signal detection means independently of each other and without interference one with another,whereby quantitative comparison of the respective signal levels detected from the first and second detectable species enables ratiometric detection of the analyte species.

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