Methods for evaluation of the effectiveness of a drug or drug candidate in treating an inflammatory bowel disease by use of a multiplexed assay kit comprising various biomarkers

Abstract

Disclosed are methods for conducting diagnostic tests for the detection of the inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis. Also described are methods for monitoring a patient by administering tests of the present invention. Also described are methods for monitoring patient's treatment by administering tests of the present invention. Also described are methods for evaluating the effectiveness of a drug or a drug candidate by administering tests of the present invention to samples from patients, animal models, and cell cultures treated with a drug or a drug candidate. Also disclosed are methods for determining the usefulness of analytes, e.g. cytokines, for acting as diagnostic and monitoring markers for inflammatory bowel disease in the various methods of the invention.

1 .- 18 . (canceled) 
     
     
         19 . A multiplexed assay kit used to evaluate the efficacy of a treatment regimen in a patient diagnosed with inflammatory bowel disease, wherein said kit comprises an assay cartridge for conducting a plurality of assays, wherein said cartridge is configured to measure a level of a plurality of biomarkers in a patient sample, said plurality of biomarkers comprises soluble TNF receptor II (sTNFRII) and MIP-1beta, and at least one additional biomarker comprising eotaxin, sIL-6R, MCP-1, RANTES, and combinations thereof. 
     
     
         20 . The kit of  claim 19  wherein the multi-well assay plate is capable of communicating with a processor, the processor configured to determine a ratio of sTNFRII to MIP-1 beta in samples from said patients versus people not diagnosed with inflammatory bowel disease. 
     
     
         21 . The kit of  claim 19 , further comprising in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said plurality of biomarkers. 
     
     
         22 . The kit of  claim 19 , wherein said kit is further configured to compare said level to a level of a normal control. 
     
     
         23 . The kit of  claim 19 , wherein said at least one additional biomarker comprises MCP-1. 
     
     
         24 . The kit of  claim 19 , wherein said kit is configured to measure said level using an immunoassay. 
     
     
         25 . The kit of  claim 19 , wherein said cartridge comprises 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 and said at least one additional biomarker in said sample. 
     
     
         26 . The kit of  claim 19 , 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. 
     
     
         27 . The kit of  claim 21 , further comprising, in said one or more vials, containers, or compartments, a set of calibrator proteins. 
     
     
         28 . The kit of  claim 19 , further comprising one or more diluents. 
     
     
         29 . The kit of  claim 21 , wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         30 . The kit of  claim 19 , wherein said kit further comprises an ECL read buffer. 
     
     
         31 . The kit of  claim 19 , further comprising capture antibodies to at least two additional biomarkers selected from the group consisting of eotaxin, sIL-6R, MCP-1, and RANTES. 
     
     
         32 . The kit of  claim 20 , wherein the processor is further configured to detect a ratio of sTNFRII to sIL-6R in samples from said patients versus people not diagnosed with inflammatory bowel disease. 
     
     
         33 . The kit of  claim 20 , wherein the processor is further configured to detect a ratio of MIP-1beta to MCP-1 in samples from said patients versus people not diagnosed with inflammatory bowel disease. 
     
     
         34 . The kit of  claim 20 , wherein the processor is further configured to detect a ratio of RANTES to MCP-1 in samples from said patients versus people not diagnosed with inflammatory bowel disease. 
     
     
         35 . The kit of  claim 19 , wherein the samples from said patients are selected from the group consisting of blood, serum, plasma, fecal, urine and combinations thereof.