Devices and methods for minimizing hook effect inteference in immunoassays

Abstract

Methods of minimizing hook effect interference in an immunoassay are disclosed. Also disclosed are reagents, kits, and immunoassay devices that may be utilized in accordance with the method.

1 . A method for detecting the presence and/or concentration of a target peptide or protein analyte in a sample, comprising the steps of:
 (a) combining, either simultaneously or wholly or partially sequentially:
 (1) a sample suspected of containing the target peptide or protein analyte; 
 (2) at least one dye capable of binding to the target peptide or protein analyte, wherein an absorption band of the at least one dye shifts upon binding to the target peptide or protein analyte; and 
 (3) at least one immunoassay reagent capable of specifically binding to the target peptide or protein analyte and capable of directly or indirectly generating a signal upon binding to the target peptide or protein analyte; 
   (b) allowing the binding of (2) and (3) to target peptide or protein analyte present in (1);   (c) determining the presence and/or concentration of the target peptide or protein analyte by detecting:
 (i) the signal generated by the immunoassay reagent; and 
 (ii) the absorption band of the at least one dye. 
   
     
     
         2 . The method of  claim 1 , wherein the at least one dye specifically binds to the target peptide or protein analyte. 
     
     
         3 . The method of  claim 1 , wherein the at least one dye non-specifically binds to the target peptide or protein analyte. 
     
     
         4 . The method of  claim 1 , wherein the at least one dye is a pH indicator. 
     
     
         5 . The method of  claim 4 , wherein the at least one dye is selected from the group comprising thymol blue, phenolphthalein, Universal pH Indicator, Coomassie Protein Assay reagent, 660 nm Protein Assay Reagent, and combinations thereof. 
     
     
         6 . The method of  claim 1 , wherein the at least one immunoassay reagent comprises a capture antibody that binds to the target analyte and a detector antibody that binds to the capture antibody. 
     
     
         7 . The method of  claim 1  wherein the signal generated by the immunoassay reagent is detected via an agglutination assay. 
     
     
         8 . The method of  claim 1 , wherein the target peptide or protein analyte is selected from the group comprising albumin, creatinine, ketone, bilirubin, urobilinogen, glycosylated hemoglobin (HbA1C), human chorionic gonadotropin (hCG), ferritin, growth hormone, prolactin, and thyroglobulin (Tg). 
     
     
         9 . The method of  claim 1 , further defined as a method of minimizing hook effect interference in an immunoassay. 
     
     
         10 . A reagent kit for use in an immunoassay, comprising:
 at least one immunoassay reagent capable of specifically binding to a target peptide or protein analyte and capable of directly or indirectly generating a signal upon binding to the target peptide or protein analyte; and   at least one dye capable of binding to the target peptide or protein analyte, wherein an absorption band of the at least one dye shifts upon binding to the target peptide or protein analyte.   
     
     
         11 . The reagent kit of  claim 10 , wherein the at least one dye is present in an assay buffer. 
     
     
         12 . The reagent kit of  claim 10 , wherein the at least one dye is in the form of a dried reagent. 
     
     
         13 . The reagent kit of  claim 10  wherein the at least one dye specifically binds to the target peptide or protein analyte. 
     
     
         14 . The reagent kit of  claim 10 , wherein the at least one dye non-specifically binds to the target peptide or protein analyte. 
     
     
         15 . The reagent kit of  claim 10  wherein the at least one dye is a pH indicator. 
     
     
         16 . The reagent kit of  claim 15 , wherein the at least one dye is selected from the group comprising thymol blue, phenolphthalein, Universal pH Indicator, Coomassie Protein Assay reagent, 660 nm Protein Assay Reagent, and combinations thereof. 
     
     
         17 . The reagent kit of  claim 10  wherein the at least one immunoassay reagent comprises a capture antibody that binds to the target analyte and a detector antibody that binds to the capture antibody. 
     
     
         18 . The reagent kit of  claim 10 , wherein the target peptide or protein analyte is selected from the group comprising albumin, creatinine, ketone, bilirubin, urobilinogen, glycosylated hemoglobin (HbA1C), human chorionic gonadotropin (hCG), ferritin, growth hormone, prolactin, and thyroglobulin (Tg). 
     
     
         19 . An immunoassay device for determining the presence and/or concentration of a target peptide or protein analyte in a sample, the immunoassay device comprising:
 at least one compartment capable of receiving a sample suspected of containing the target peptide or protein analyte, the at least one compartment comprising:
 at least one immunoassay reagent capable of specifically binding to a target peptide or protein analyte and capable of directly or indirectly generating a signal upon binding to the target peptide or protein analyte; and 
 at least one dye capable of binding to the target peptide or protein analyte, wherein an absorption band of the at least one dye shifts upon binding to the target peptide or protein analyte. 
   
     
     
         20 . The immunoassay device of  claim 19 , further defined as comprising at least two compartments capable of being in fluidic communication with one another, wherein the at least one immunoassay reagent and the at least one dye are disposed in the same compartment.