Biomarkers of therapeutic responsiveness

Abstract

The present invention relates to methods of diagnosing a kidney disorder in a patient, as well as methods of monitoring the progression of a kidney disorder and/or methods of monitoring a treatment protocol of a therapeutic agent or a therapeutic regimen. The invention also relates to assay methods used in connection with the diagnostic methods described herein.

1 .- 16 . (canceled) 
     
     
         17 . A kit for the analysis of a renal cell carcinoma (RCC) cancer panel comprising an assay cartridge for conducting a plurality of assays, said cartridge comprising a flow cell having an inlet, an outlet or a detection chamber, said inlet, detecting chamber, or outlet defining a flow path through said flow cell, said detection chamber comprising a plurality of binding domains, at least two of said domains comprising a set of capture antibodies to a pair of human biomarkers, said set comprising one or both of a first capture antibody specific for a total form of said human biomarker and a second capture antibody specific for a phosphorylated form of said human biomarker of one or more of the following groups,
 wherein in a first group an increase in the concentration of the biomarker from a first level in a treated, resistant RCC tumor indicates a responsive RCC tumor, the first group of biomarkers selected from the group consisting of total Akt, AMPKa1 (p174), and phosphorylated STAT,   wherein in a second group a decrease in the concentration of the biomarker from a first level in an untreated RCC tumor indicates a non-responsive RCC tumor, the second group of biomarkers selected from total Akt and APMKa1 (p174),   wherein in a third group an increase in the concentration of the biomarker from a first level in an untreated RCC tumor indicates a non-responsive RCC tumor, the third group selected from phosphorylated Erk1/2 and phosphorylated Gsk3b,   wherein in a fourth group a decrease in the concentration of the biomarker from a first level in a treated, resistant RCC tumor indicates a responsive RCC tumor, the fourth group of biomarkers selected from the group consisting of phosphorylated Erk1/2, total VEGF, total PIGF, total VEGFR-1, and total p21,   wherein in a fifth group an increase in the concentration of the biomarker from a first level in an untreated RCC tumor indicates a responsive RCC tumor, the fifth group consisting of total Met,   wherein in a sixth group an increase in the concentration of the biomarker from a first level in a treated, responsive RCC tumor indicates a non-responsive RCC tumor, the sixth group of biomarkers is phosphorylated Erk1/2, total VEGF, total PIGF, total VEGFR-1, and total p21, and   wherein in a seventh group a decrease in the concentration of the biomarker from a first level in an untreated or a treated, responsive RCC tumor indicates a non-responsive RCC tumor, the seventh group of biomarkers selected from the group consisting of total Akt, and AMPKa1 (p174),   and said cartridge further comprises, in one or more compartments, a set of labeled detection antibodies specific for said human biomarkers.   
     
     
         18 . The kit of  claim 17  wherein said kit further comprises one or more additional assay reagents used in said assay, said one or more additional assay reagents provided in one or more compartments of said, wherein said one or more additional assay reagents are selected from the group consisting of a set of calibrator proteins, one or more diluents, electrochemiluminescence (ECL) read buffer, and a combination thereof. 
     
     
         19 . A kit for the analysis of a renal cell carcinoma (RCC) cancer panel comprising
 (a) a multi-well assay plate comprising a plurality of wells, each well comprising a plurality of discrete binding domains, at least two of said domains comprising a set of capture antibodies to a pair of human biomarkers, said set comprising one or both of a first capture antibody specific for a total form of said human biomarker and a second capture antibody specific for a phosphorylated form of said human biomarker of one or more of the following groups,   wherein in a first group an increase in the concentration of the biomarker from a first level in a treated, resistant RCC tumor indicates a responsive RCC tumor, the first group of biomarkers selected from the group consisting of total Akt, AMPKa1 (p174), and phosphorylated STAT,   wherein in a second group a decrease in the concentration of the biomarker from a first level in an untreated RCC tumor indicates a non-responsive RCC tumor, the second group of biomarkers selected from total Akt and APMKa1 (p174),   wherein in a third group an increase in the concentration of the biomarker from a first level in an untreated RCC tumor indicates a non-responsive RCC tumor, the third group selected from phosphorylated Erk1/2 and phosphorylated Gsk3b,   wherein in a fourth group an decrease in the concentration of the biomarker from a first level in a treated, resistant RCC tumor indicates a responsive RCC tumor, the fourth group of biomarkers selected from the group consisting of phosphorylated Erk1/2, total VEGF, total PIGF, total VEGFR-1, and total p21,   wherein in a fifth group an increase in the concentration of the biomarker from a first level in an untreated RCC tumor indicates a responsive RCC tumor, the fifth group consisting of total Met,   wherein in a sixth group an increase in the concentration of the biomarker from a first level in a treated, responsive RCC tumor indicates a non-responsive RCC tumor, the sixth group of biomarkers is phosphorylated Erk1/2, total VEGF, total PIGF, total VEGFR-1, and total p21, and   wherein in a seventh group a decrease in the concentration of the biomarker from a first level in an untreated or a treated, responsive RCC tumor indicates a non-responsive RCC tumor, the seventh group of biomarkers selected from the group consisting of total Akt, and AMPKa1 (p174);   (b) in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said human biomarkers; and   (c) in one or more vials, containers, or compartments, a set of calibrator proteins.   
     
     
         20 . The kit of  claim 19  wherein said kit further comprises one or more diluents. 
     
     
         21 . The kit of  claim 19  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         22 . The kit of  claim 19  wherein said kit further comprises an ECL read buffer. 
     
     
         23 . The kit of  claim 19  wherein said discrete binding domains are positioned on an electrode within said well. 
     
     
         24 . The kit of  claim 19  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         25 . The kit of  claim 19  wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins. 
     
     
         26 . The kit of  claim 17  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         27 . The kit of  claim 18  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         28 . The kit of  claim 18  wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins. 
     
     
         29 . The kit of  claim 17 , wherein said plurality of binding domains are particles. 
     
     
         30 . The kit of  claim 19 , wherein said plurality of binding domains are particles. 
     
     
         31 . A kit for the analysis of a renal cell carcinoma (RCC) cancer panel comprising an assay cartridge for conducting a plurality of assays, said cartridge comprising a flow cell having an inlet, an outlet or a detection chamber, said inlet, detecting chamber, or outlet defining a flow path through said flow cell, said detection chamber comprising a plurality of binding domains, at least two of said domains comprising a set of capture antibodies to a pair of human biomarkers, said set comprising a first capture antibody specific for a total form of said human biomarker and a second capture antibody specific for a phosphorylated form of said human biomarker, wherein the human biomarkers are selected from the group consisting of Akt, Erk1/2, Met, GSK3b, Hif1a, p21, AMPKa1, VEGF, P1GF, VEGFR-1/Flt-1, STAT3 and a combination thereof, wherein in a first group a decrease in the concentration of the biomarker from a first level indicates a non-responsive RCC tumor, the first group of biomarkers selected from the group consisting of the total form of Akt, the total form of Erk1/2, the total form of Met, the total form of AMPKa1, the total form of Hif1a, the phosphorylated form of STAT3, and combinations thereof, and wherein in a second group an increase in the concentration of the biomarker from a first level indicates a non-responsive RCC tumor, the second group of biomarkers selected from the group consisting of the phosphorylated form of Akt, the phosphorylated form of Erk1/2, the phosphorylated form of Met, the total form of Gsk3b, the phosphorylated form of Gsk3b, the total form of VEGF, the total form of P1GF, the total form of VEGFR-1, the total form of p21, and combinations thereof, and said cartridge further comprises, in one or more compartments, a set of labeled detection antibodies specific for said human biomarkers. 
     
     
         32 . The kit of  claim 31  wherein said kit further comprises one or more additional assay reagents used in said assay, said one or more additional assay reagents provided in one or more compartments of said, wherein said one or more additional assay reagents are selected from the group consisting of a set of calibrator proteins, one or more diluents, electrochemiluminescence (ECL) read buffer, and a combination thereof. 
     
     
         33 . The kit of  claim 31  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         34 . The kit of  claim 32  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         35 . The kit of  claim 32 , wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins. 
     
     
         36 . The kit of  claim 31 , wherein said plurality of binding domains are particles. 
     
     
         37 . A kit for the analysis of a renal cell carcinoma (RCC) cancer panel comprising
 (a) a multi-well assay plate comprising a plurality of wells, each well comprising a plurality of discrete binding domains, at least two of said domains comprising a set of capture antibodies to a pair of human biomarkers, said set comprising a first capture antibody specific for a total form of said human biomarker and a second capture antibody specific for a phosphorylated form of said human biomarker, wherein the human biomarkers are selected from the group consisting of: Akt, Erk1/2, Met, GSK3b, Hif1a, p21, AMPKa1, VEGF, P1GF, VEGFR-1/Flt-1, STAT3 and a combination thereof, wherein in a first group a decrease in the concentration of the biomarker from a first level indicates a non-responsive RCC tumor, the first group of biomarkers selected from the group consisting of the total form of Akt, the total form of Erk1/2, the total form of Met, the total form of AMPKa1, the total form of Hif1a, the phosphorylated form of STAT3, and combinations thereof, and wherein in a second group an increase in the concentration of the biomarker from a first level indicates a non-responsive RCC tumor, the second group of biomarkers selected from the group consisting of the phosphorylated form of Akt, the phosphorylated form of Erk1/2, the phosphorylated form of Met, the total form of Gsk3b, the phosphorylated form of Gsk3b, the total form of VEGF, the total form of P1GF, the total form of VEGFR-1, the total form of p21, and combinations thereof;   (b) in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said human biomarkers; and   (c) in one or more vials, containers, or compartments, a set of calibrator proteins.   
     
     
         38 . The kit of  claim 37  wherein said kit further comprises one or more diluents. 
     
     
         39 . The kit of  claim 37  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         40 . The kit of  claim 37  wherein said kit further comprises an ECL read buffer. 
     
     
         41 . The kit of  claim 37  wherein said discrete binding domains are positioned on an electrode within said well. 
     
     
         42 . The kit of  claim 37  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         43 . The kit of  claim 37  wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins.