Lung cancer biomarkers

Abstract

The present invention relates to methods of diagnosing lung cancer in a patient, as well as methods of monitoring the progression of lung cancer 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.

What is claimed is: 
     
         1 . A method for evaluating the efficacy of a treatment regimen in a patient diagnosed with lung cancer, said method comprising
 (a) obtaining a test sample from a patient undergoing said treatment regimen for lung cancer;   (b) measuring a level of a biomarker in said test sample, wherein said biomarker comprises MDC, NME-2, KGF, PIGF, Flt-3L, HGF, MCP1, SAT-1, MIP-1-b, GCLM, OPG, TNF RII, VEGF-D, ITAC, MMP-10, GPI, PPP2R4, AKR1B1, AmylA, MIP-1b, P-Cadherin, EPO, MMP-2, EGFR, MMP-3, ErbB2, cytokeratin-19, E-cadherin, IL-6, osteopontin, cKit, uPA, NSE, cMET, MDC, Flt-1, CEA, cytokeratin-8, KGF, S100A6, IL2-R, and combinations thereof;   (c) comparing said level to a normal control level of said biomarker; and   (d) evaluating from said comparing step (c) whether said patient is responsive to said treatment regimen.   
     
     
         2 . The method of  claim 1  wherein said measuring step comprises conducting a multiplexed assay measurement of a plurality of said biomarkers in said test sample, wherein said multiplexed assay measurement is conducted using one reaction volume comprising said test sample. 
     
     
         3 . The method of  claim 1  wherein said measuring step(s) are conducted on a single sample. 
     
     
         4 . The method of  claim 1  wherein said measuring step(s) are conducted in a single assay chamber. 
     
     
         5 . The method of  claim 4  wherein said assay chamber is a single well of an assay plate. 
     
     
         6 . The method of  claim 4  wherein said assay chamber is a cartridge. 
     
     
         7 . The method of  claim 1  wherein said sample is selected from the group consisting of blood, peripheral blood mononuclear cells (PBMC), isolated blood cells, urine, serum and plasma. 
     
     
         8 . The method of  claim 1  wherein said sample is selected from serum or plasma. 
     
     
         9 . The method of  claim 1  wherein said sample is serum. 
     
     
         10 . The method of  claim 1  wherein said sample is plasma. 
     
     
         11 . The method of  claim 1  wherein said level(s) are measured using an immunoassay. 
     
     
         12 . A multiplexed assay kit used to evaluate the efficacy of a treatment regimen in a patient diagnosed with lung cancer, said kit is configured to measure a level of a plurality of biomarkers in a patient sample, said plurality of biomarkers comprises MDC, NME-2, KGF, PIGF, Flt-3L, HGF, MCP1, SAT-1, MIP-1-b, GCLM, OPG, TNF RII, VEGF-D, ITAC, MMP-10, GPI, PPP2R4, AKR1B1, AmylA, MIP-1b, P-Cadherin, EPO, MMP-2, EGFR, MMP-3, ErbB2, cytokeratin-19, E-cadherin, IL-6, osteopontin, cKit, uPA, NSE, cMET, MDC, Flt-1, CEA, cytokeratin-8, KGF, S100A6, IL2-R, and combinations thereof. 
     
     
         13 . The kit of  claim 12  wherein said kit is further configured to compare said level to a level of a normal control. 
     
     
         14 . The kit of  claim 12  wherein said kit is configured to measure said level using an immunoassay. 
     
     
         15 . The kit of  claim 14  wherein said kit comprises a multi-well assay plate including a plurality of assay wells used in an assay conducted in said kit, said plurality of assay wells configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         16 . The kit of  claim 15  wherein a well of said assay plate comprises a plurality of assay domains, at least two of said assay domains comprising reagents for measuring different biomarkers. 
     
     
         17 . The kit of  claim 14  wherein said kit comprises 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 configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         18 . The kit of  claim 12  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 vials, containers, or compartments of said kit. 
     
     
         19 . A kit for the analysis of a lung cancer panel comprising
 (a) a multi-well assay plate comprising a plurality of wells, each well comprising at least four discrete binding domains to which capture antibodies to the following human analytes are bound: MDC, NME-2, KGF, PIGF, Flt-3L, HGF, MCP1, SAT-1, MIP-1-b, GCLM, OPG, TNF RII, VEGF-D, ITAC, MMP-10, GPI, PPP2R4, AKR1B1, AmylA, MIP-1b, P-Cadherin, EPO, MMP-2, EGFR, MMP-3, ErbB2, cytokeratin-19, E-cadherin, IL-6, osteopontin, cKit, uPA, NSE, cMET, MDC, Flt-1, CEA, cytokeratin-8, KGF, S100A6, IL2-R, and combinations thereof;   (b) in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said human analytes; 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 21  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 . A multiplexed assay kit used to evaluate the efficacy of a treatment regimen in a patient diagnosed with lung cancer, said kit is configured to measure a level of a plurality of biomarkers in a patient serum sample, said plurality of biomarkers comprises Flt-3L, EGFR, MMP-3, and NME-2, and combinations thereof. 
     
     
         27 . The kit of  claim 26  wherein said kit is further configured to compare said level to a level of a normal control. 
     
     
         28 . The kit of  claim 26  wherein said kit is configured to measure said level using an immunoassay. 
     
     
         29 . The kit of  claim 28  wherein said kit comprises a multi-well assay plate including a plurality of assay wells used in an assay conducted in said kit, said plurality of assay wells configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         30 . The kit of  claim 29  wherein a well of said assay plate comprises a plurality of assay domains, at least two of said assay domains comprising reagents for measuring different biomarkers. 
     
     
         31 . The kit of  claim 28  wherein said kit comprises 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 configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         32 . The kit of  claim 26  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 vials, containers, or compartments of said kit. 
     
     
         33 . A kit for the analysis of a lung cancer panel comprising
 (a) a multi-well assay plate comprising a plurality of wells, each well comprising at least four discrete binding domains to which capture antibodies to the following human serum analytes are bound: Flt-3L, EGFR, MMP-3, and NME-2, and combinations thereof;   (b) in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said human analytes; and   (c) in one or more vials, containers, or compartments, a set of calibrator proteins.   
     
     
         34 . The kit of  claim 33  wherein said kit further comprises one or more diluents. 
     
     
         35 . The kit of  claim 33  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         36 . The kit of  claim 35  wherein said kit further comprises an ECL read buffer. 
     
     
         37 . The kit of  claim 33  wherein said discrete binding domains are positioned on an electrode within said well. 
     
     
         38 . The kit of  claim 33  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         39 . The kit of  claim 33  wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins. 
     
     
         40 . A multiplexed assay kit used to evaluate the efficacy of a treatment regimen in a patient diagnosed with lung cancer, said kit is configured to measure a level of a plurality of biomarkers in a patient plasma sample, said plurality of biomarkers comprises Flt-3L, cytokeratin-19, Flt-1, KGF, and HGF, and combinations thereof. 
     
     
         41 . The kit of  claim 40  wherein said kit is further configured to compare said level to a level of a normal control. 
     
     
         42 . The kit of  claim 40  wherein said kit is configured to measure said level using an immunoassay. 
     
     
         43 . The kit of  claim 42  wherein said kit comprises a multi-well assay plate including a plurality of assay wells used in an assay conducted in said kit, said plurality of assay wells configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         44 . The kit of  claim 43  wherein a well of said assay plate comprises a plurality of assay domains, at least two of said assay domains comprising reagents for measuring different biomarkers. 
     
     
         45 . The kit of  claim 43  wherein said kit comprises 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 configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         46 . The kit of  claim 40  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 vials, containers, or compartments of said kit. 
     
     
         47 . A kit for the analysis of a lung cancer panel comprising
 (a) a multi-well assay plate comprising a plurality of wells, each well comprising at least four discrete binding domains to which capture antibodies to the following human plasma analytes are bound: Flt-3L, cytokeratin-19, Flt-1, KGF, and HGF, and combinations thereof;   (b) in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said human analytes; and   (c) in one or more vials, containers, or compartments, a set of calibrator proteins.   
     
     
         48 . The kit of  claim 47  wherein said kit further comprises one or more diluents. 
     
     
         49 . The kit of  claim 47  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         50 . The kit of  claim 49  wherein said kit further comprises an ECL read buffer. 
     
     
         51 . The kit of  claim 47  wherein said discrete binding domains are positioned on an electrode within said well. 
     
     
         52 . The kit of  claim 47  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         53 . The kit of  claim 47  wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins.