Protein biomarker and uses thereof

Abstract

The present invention is directed to methods of identifying and treating a human subject harboring a tumor or other disease comprising assessing HRG gene expression at a protein level in the human subject and administering a treatment comprising an anti-HER3 antibody to the human subject whose HRG gene expression at a protein level is assessed as high. The present invention is also directed to methods of identifying a human subject harboring a tumor or other disease comprising assessing HRG gene expression at a protein level in the human subject and withholding a treatment comprising an anti-HER3 antibody to the human subject whose HRG gene expression at a protein level is assessed as low. The invention is also directed to methods of performing an ELISA, including sequential steps of contacting a solid surface with a plurality of solutions each comprising in turn a capture antibody, a blocking agent, a sample suspected of containing an analyte, a detection antibody and an enzyme conjugate, in which the solid surface is subjected to a wash process after each sequential step.

1 - 74 . (canceled) 
     
     
         75 . A method of treating a human subject harboring a locally advanced or metastatic non-small cell lung cancer (NSCLC) tumor comprising administering a treatment comprising an anti-HER3 antibody to a human subject diagnosed with a locally advanced or metastatic NSCLC whose HRG gene expression at a protein level is assessed as high. 
     
     
         76 . The method of  claim 75  in which the HRG gene expression at a protein level is assessed as high if a protein concentration value is observed, which is above a predetermined threshold, from a biological sample taken from the subject diagnosed with a locally advanced or metastatic NSCLC. 
     
     
         77 . The method of  claim 76  in which the predetermined threshold is statistically determined, refined, adjusted and/or confirmed through, on, or based on, randomized clinical data and optionally non-clinical data. 
     
     
         78 . The method of  claim 76  in which the predetermined threshold value is selected from the group consisting of 0 pg/mL, about 980 pg/mL, about 1622 pg/mL, about 2000 pg/mL, about 3000 pg/mL, about 4000 pg/mL, about 5000 pg/mL, about 6000 pg/mL, about 7000 pg/mL, about 8000 pg/mL, about 9000 pg/mL, about 10000 pg/mL, about 11000 pg/mL, about 12000 pg/mL, about 13000 pg/mL, about 14000 pg/mL, about 15000 pg/mL, about 16000 pg/mL, about 17000 pg/mL, about 18000 pg/mL, about 19000 pg/mL, about 20000 pg/mL, about 22000 pg/mL, about 24000 pg/mL, about 26000 pg/mL, about 28000 pg/mL, about 30000 pg/mL, about 35000 pg/mL, about 40000 pg/mL, about 50000 pg/mL, about 60000 pg/mL, about 70000 pg/mL, about 80000 pg/Lm, about 90000 pg/mL and about 100000 pg/mL. 
     
     
         79 . The method of  claim 75 , wherein the subject harbors wild-type EGFR. 
     
     
         80 . The method of  claim 75 , wherein the tumor has progressed on at least one prior systemic therapy. 
     
     
         81 . The method of  claim 75 , further comprising assessing gene expression at a protein level in the human subject diagnosed with the locally advanced or metastatic NSCLC, wherein the HRG gene expression at a protein level is assessed using ELISA or immunohistochemistry techniques. 
     
     
         82 . The method of  claim 76  in which the biological sample comprises a whole blood or serum sample. 
     
     
         83 . The method of  claim 75  in which the anti-HER3 antibody is selected from the group consisting of patritumab, duligotumab (MEHD-7945A), seribantumab (MM-121), MM-111, LJM716, RG-7116, tri-specific anti-EGFR/ERBB3 zybody, huHER3-8, and a derivative or fragment of any of these. 
     
     
         84 . The method of  claim 75  in which the treatment comprises an anti-HER3 antibody in combination with one or more of (i) a HER inhibitor, (ii) a chemotherapy, (iii) radiation and (iv) an other targeted agent. 
     
     
         85 . The method of  claim 84 , wherein the HER inhibitor is selected from the group consisting of trastuzumab, T-DM1, lapatinib, pertuzumab, cetuximab, panitumumab gefitinib, afatinib, dacomitinib, KD-019 and erlotinib. 
     
     
         86 . The method of  claim 84 , wherein the chemotherapy is selected from the group consisting of cisplatin, carboplatin, gemcitabine, pemetrexed, irinotecan, 5-fluoruracil, paclitaxel, docetaxel, and capecitabine. 
     
     
         87 . The method of  claim 75 , wherein the HRG expression is assessed using an FDA-approved test. 
     
     
         88 . A method of treating a human subject harboring a locally advanced or metastatic non-small cell lung cancer (NSCLC) tumor comprising withholding a treatment comprising an anti-HER3 antibody from a human subject diagnosed with a locally advanced or metastatic NSCLC whose HRG gene expression at a protein level is assessed as low. 
     
     
         89 . The method of  claim 88  in which the HRG gene expression at a protein level is assessed as low if a protein concentration value is observed, which is at or below a predetermined threshold, from a biological sample taken from the subject diagnosed with a locally advanced or metastatic NSCLC. 
     
     
         90 . The method of  claim 89  in which the predetermined threshold is statistically determined, refined, adjusted and/or confirmed through, on, or based on, randomized clinical data and optionally non-clinical data. 
     
     
         91 . The method of  claim 90  in which the predetermined threshold value is selected from the group consisting of 0 pg/mL, about 980 pg/mL, about 1000 pg/mL, about 1622 pg/mL, about 2000 pg/mL, about 3000 pg/mL, about 4000 pg/mL, about 5000 pg/mL, about 6000 pg/mL, about 7000 pg/mL, about 8000 pg/mL, about 9000 pg/mL, about 10000 pg/mL, about 11000 pg/mL, about 12000 pg/mL, about 13000 pg/mL, about 14000 pg/mL, about 15000 pg/mL, about 16000 pg/mL, about 17000 pg/mL, about 18000 pg/mL, about 19000 pg/mL, about 20000 pg/mL, about 22000 pg/mL, about 24000 pg/mL, about 26000 pg/mL, about 28000 pg/mL, about 30000 pg/mL, about 35000 pg/mL, about 40000 pg/mL, about 50000 pg/mL, about 60000 pg/mL, about 70000 pg/mL, about 80000 pg/mL, about 90000 pg/mL and about 100000 pg/mL. 
     
     
         92 . The method of  claim 88 , wherein the subject harbors wild-type EGFR. 
     
     
         93 . The method of  claim 88 , wherein the tumor has progressed on at least one prior systemic therapy. 
     
     
         94 . The method of  claim 89  in which the biological sample comprises a whole blood or serum sample. 
     
     
         95 . The method of  claim 88  in which the treatment withheld comprises an anti-HER3 antibody in combination with one or more of (i) a HER inhibitor, (ii) a chemotherapy, (iii) radiation and (iv) an other targeted agent. 
     
     
         96 . A method of receiving or undergoing a treatment for a locally advanced or metastatic non-small cell lung cancer (NSCLC) tumor or abstaining therefrom comprising:
 providing an autologous tissue sample or consenting to a taking of same to facilitate an assessment of HRG gene expression at a protein level in a human subject diagnosed with a locally advanced or metastatic NSCLC; and   receiving or undergoing a treatment comprising an anti-HER3 antibody if the HRG gene expression at a protein level is assessed as high, or   abstaining from a treatment comprising an anti-HER3 antibody if the HRG gene expression at a protein level is assessed as low.   
     
     
         97 . The method of  claim 96  in which (i) the HRG gene expression at a protein level is assessed as high if a protein concentration value is observed, which is above a predetermined threshold, from a biological sample taken from the subject diagnosed with a locally advanced or metastatic NSCLC or (ii) the HRG gene expression at a protein level is assessed as low if a protein concentration value is observed, which is at or below the predetermined threshold, from the biological sample taken from the subject diagnosed with a locally advanced or metastatic NSCLC. 
     
     
         98 . The method of  claim 97  in which the predetermined threshold is statistically determined, refined, adjusted and/or confirmed through, on, or based on, randomized clinical data and optionally non-clinical data. 
     
     
         99 . The method of  claim 98  in which the predetermined threshold value is selected from the group consisting of 0 pg/mL, about 980 pg/mL, about 1622 pg/mL, about 2000 pg/mL, about 3000 pg/mL, about 4000 pg/mL, about 5000 pg/mL, about 6000 pg/mL, about 7000 pg/mL, about 8000 pg/mL, about 9000 pg/mL, about 10000 pg/mL, about 11000 pg/mL, about 12000 pg/mL, about 13000 pg/mL, about 14000 pg/mL, about 15000 pg/mL, about 16000 pg/mL, about 17000 pg/mL, about 18000 pg/mL, about 19000 pg/mL, about 20000 pg/mL, about 22000 pg/mL, about 24000 pg/mL, about 26000 pg/mL, about 28000 pg/mL, about 30000 pg/mL, about 35000 pg/mL, about 40000 pg/mL, about 50000 pg/mL, about 60000 pg/mL, about 70000 pg/mL, about 80000 pg/Lm, about 90000 pg/mL and about 100000 pg/mL. 
     
     
         100 . A method of electing a treatment for a locally advanced or metastatic non-small cell lung cancer (NSCLC) tumor comprising:
 receiving an assessment of HRG gene expression at a protein level in a human subject diagnosed with a locally advanced or metastatic NSCLC; and   electing to withhold or abstain from a treatment comprising an anti-HER3 antibody if HRG gene expression at a protein level is assessed as low, or   electing to receive or undergo a treatment comprising an anti-HER3 antibody if HRG gene expression at a protein level is assessed as high.   
     
     
         101 . The method of  claim 100  in which (i) the HRG gene expression at a protein level is assessed as high if a protein concentration value is observed, which is above a predetermined threshold, from a biological sample taken from the subject diagnosed with a locally advanced or metastatic NSCLC or (ii) the HRG gene expression at a protein level is assessed as low if a protein concentration value is observed, which is at or below the predetermined threshold, from the biological sample taken from the subject diagnosed with a locally advanced or metastatic NSCLC. 
     
     
         102 . The method of  claim 101  in which the predetermined threshold is statistically determined, refined, adjusted and/or confirmed through, on, or based on, randomized clinical data and optionally non-clinical data. 
     
     
         103 . The method of  claim 102  in which the predetermined threshold value is selected from the group consisting of 0 pg/mL, about 980 pg/mL, about 1622 pg/mL, about 2000 pg/mL, about 3000 pg/mL, about 4000 pg/mL, about 5000 pg/mL, about 6000 pg/mL, about 7000 pg/mL, about 8000 pg/mL, about 9000 pg/mL, about 10000 pg/mL, about 11000 pg/mL, about 12000 pg/mL, about 13000 pg/mL, about 14000 pg/mL, about 15000 pg/mL, about 16000 pg/mL, about 17000 pg/mL, about 18000 pg/mL, about 19000 pg/mL, about 20000 pg/mL, about 22000 pg/mL, about 24000 pg/mL, about 26000 pg/mL, about 28000 pg/mL, about 30000 pg/mL, about 35000 pg/mL, about 40000 pg/mL, about 50000 pg/mL, about 60000 pg/mL, about 70000 pg/mL, about 80000 pg/Lm, about 90000 pg/mL and about 100000 pg/mL.