Assay to detect human dpp-4

Abstract

This disclosure provides a robust, sensitive, and specific assay for the detection and measurement of DPP-4 levels in samples obtained from human patients. The disclosure further provides novel anti-DPP-4 monoclonal antibodies that recognize human DPP-4, and assay kits comprising one or more of these antibodies.

1 .- 54 . (canceled) 
     
     
         55 . A kit for measuring dipeptidyl peptidase-4 (DPP-4) levels in a sample, comprising one or more isolated antibodies that binds to human dipeptidyl peptidase-4 (DPP-4) and instructions for use thereof, wherein each of the one or more isolated antibodies comprises a heavy chain variable domain (VH) with three heavy chain complementarity determining regions (CDRs) VHCDR1, VHCDR2 and VHCDR3, and a light chain variable domain (VL) with three light chain CDRs VLCDR1, VLCDR2, and VLCDR3, wherein the six CDRs of each isolated antibody are identical to the six CDRs of mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the American Type Culture Collection (ATCC) under Deposit No. PTA-121870, mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871, rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872, or rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873. 
     
     
         56 . The kit of  claim 55 , which comprises two isolated antibodies. 
     
     
         57 . The kit of  claim 56 , wherein one isolated antibody is a capture antibody and the other isolated antibody is a detection antibody. 
     
     
         58 . The kit of  claim 57 , wherein:
 (a) the capture antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870 and the detection antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872;   (b) the capture antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870 and the detection antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873;   (c) the capture antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871 and the detection antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872;   (d) the capture antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871 and the detection antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873;   (e) the capture antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872 and the detection antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870;   (f) the capture antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872 and the detection antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871;   (g) the capture antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873 and the detection antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870; or   (h) the capture antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873 and the detection antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871.   
     
     
         59 . The kit of  claim 58 , wherein the detection antibody comprises a detectable label. 
     
     
         60 . The kit of  claim 59 , wherein the detectable label is biotin. 
     
     
         61 . The kit of  claim 59 , wherein the detectable label is a ruthenium chelate. 
     
     
         62 . The kit of  claim 55 , further comprising a solid support and detection reagents. 
     
     
         63 . A method of measuring the amount of dipeptidyl peptidase-4 (DPP-4) in a sample obtained from a subject comprising assaying the sample using the kit of  claim 55 . 
     
     
         64 . The method of  claim 63 , wherein the sample is whole blood, serum, plasma, saliva, urine, sputum, bronchoalveolar lavage fluid, lung epithelial cells, nasal polyps, or skin. 
     
     
         65 . The method of  claim 63 , wherein the subject has a disease or condition selected from an IL-13-mediated disease or disorder, a pulmonary disease or disorder, and a chronic inflammatory skin disease or disorder. 
     
     
         66 . The method of  claim 65 , wherein the disease or condition is selected from asthma, atopic asthma, corticosteroid naïve asthma, chronic asthma, corticosteroid resistant asthma, corticosteroid refractory asthma, asthma due to smoking, asthma uncontrolled on corticosteroids, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), ulcerative colitis (UC), atopic dermatitis (AD), allergic rhinitis, chronic rhinosinusitis, skin fibrosis, allergic contact dermatitis, eczema, and psoriasis. 
     
     
         67 . The method of  claim 63 , wherein the sample is assayed using a sandwich immunoassay comprising a capture antibody and a detection antibody. 
     
     
         68 . The method of  claim 67 , wherein the sandwich immunoassay comprises:
 (a) attaching a capture antibody to a solid support;   (b) applying the sample obtained from a subject or a control sample to the solid support under conditions sufficient to allow DPP-4, if present in the sample, to bind to the capture antibody;   (c) contacting the solid support with the detection antibody under conditions sufficient to allow binding of the detection antibody to DPP-4 already bound to the capture antibody; and   (d) measuring the amount of detection antibody bound to DPP-4.   
     
     
         69 . The method of  claim 67 , wherein:
 (a) the capture antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870 and the detection antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872;   (b) the capture antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870 and the detection antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873;   (c) the capture antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871 and the detection antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872;   (d) the capture antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871 and the detection antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873;   (e) the capture antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872 and the detection antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870;   (f) the capture antibody is rat monoclonal antibody R11A2.15 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121872 and the detection antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871;   (g) the capture antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873 and the detection antibody is mouse monoclonal antibody m3B7.6 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121870; or   (h) the capture antibody is rat monoclonal antibody R11A9.11 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121873 and the detection antibody is mouse monoclonal antibody m5B7.7 produced from a hybridoma deposited at the ATCC under Deposit No. PTA-121871.