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Compensation for variability in specific binding in quantitative assays

  • US 20030162236A1
  • Filed: 03/26/2001
  • Published: 08/28/2003
  • Est. Priority Date: 03/26/2001
  • Status: Abandoned Application
First Claim
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1. A method for quantitatively measuring the amount of an analyte of interest in a fluid sample, comprising:

  • a) providing a membrane strip comprising an application point, a contact region, a sample capture zone and a control capture zone, wherein the contact region is between the application point and the sample capture zone and the sample capture zone is between the contact region and the control capture zone;

    b) contacting the application point of the membrane strip with the fluid sample to be assayed for the analyte of interest;

    c) maintaining the membrane strip under conditions which allow fluid to transport analyte of interest in the fluid sample by capillary action through the strip to and through the contact region, the contact region having a population of analyte-binding particles immobilized therein, wherein the analyte-binding particles are coated with an analyte-binding agent;

    d) further maintaining the membrane strip under conditions which allow analyte of interest, if present in the sample, to bind to analyte-binding particles, thereby generating contacted analyte-binding particles;

    allow the fluid in the sample to mobilize and transport contacted analyte-binding particles by capillary action through the strip to and through the sample capture zone, the sample capture zone having a sample capture reagent immobilized thereon; and

    allow contacted analyte-binding particles to bind to the sample capture reagent;

    e) further maintaining the membrane strip under conditions which allow the fluid in the sample to transport contacted analyte-binding particles by capillary action through the strip to and through the control capture zone, the control capture zone having a control capture reagent immobilized thereon; and

    allow contacted analyte-binding particles to bind to the control capture reagent;

    f) further maintaining the membrane strip under conditions which allow the fluid in the sample to transport any contacted analyte-binding particles not bound to the sample capture reagent or to the control capture reagent by capillary action beyond the control capture zone;

    g) determining the amount of contacted analyte-binding particles in the sample capture zone and the amount of contacted analyte-binding particles in the control capture zone;

    h) determining a corrected analyte-binding particle amount from the amount of analyte-binding particles in the sample capture zone and the amount of analyte-binding particles in the control capture zone, wherein the amount of analyte of interest in the fluid sample is directly related to the corrected analyte-binding particle amount.

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