US2010143954A1PendingUtilityA1

Galectin-3 Immunoassay

Assignee: BG MEDICINE INCPriority: Oct 29, 2008Filed: Oct 29, 2009Published: Jun 10, 2010
Est. expiryOct 29, 2028(~2.3 yrs left)· nominal 20-yr term from priority
G01N 2333/4724G01N 33/6893G01N 2800/325G01N 33/68G01N 33/577C07K 16/2851G01N 33/53C07K 16/18
60
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Claims

Abstract

The present invention relates to methods and compositions for specifically and quantitatively detecting galectin-3 in a sample. Embodiments of the invention include a detection assay in which a capture binding moiety and a labeled binding moiety specifically recognize non-overlapping epitopes on the N-terminus of galectin-3. Further embodiments are directed to a method for establishing ranges of galectin-3 concentrations indicative of the presence and severity of heart failure in a subject and a method for predicting the clinical outcome of a subject based upon galectin-3 concentration.

Claims

exact text as granted — not AI-modified
1 . A kit for detecting the concentration of galectin-3 in a sample, the kit comprising
 first and second binding moieties, one being labeled with a detectable label, each binding respectively to spaced-apart epitopes on the N-terminal portion of galectin-3, which comprises the sequence of amino acids:   
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                 
                 
               
                     
                   MADNFSLHDA LSGSGNPNPQ GWPGAWGNQP AGAGGYPGAS 
                 
                     
                     
                 
                     
                   YPGAYPGQAP PGAYPGQAPP GAYPGAPGAY PGAPAPGVYP 
                 
                     
                     
                 
                     
                   GPPSGPGAYP SSGQPSATGA YPATGPYGAP AGP. 
                 
             
                
               
            
             
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The kit of  claim 1  wherein the detectable label is an enzyme or a fluorophore. 
     
     
         3 . The kit of  claim 1  wherein the binding moieties respectively bind to an epitope defined by at least a portion of a galectin-3 amino acid sequence selected from the group consisting of: 
       
         
           
                 
                 
               
                   MADNFSLHDALS, 
                   (amino acids 1-12 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                     
                 
                   MADNFSLHDALSGS, 
                   (amino acids 1-14 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                     
                 
                   GNPNPQGWPGA, 
                   (amino acids 15-25 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                     
                 
                   WGNQPAGAGG, 
                   (amino acids 26-35 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                     
                 
                   YPGQAPPGAYPGQAPPGA, 
                   (amino acids 45-62 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                     
                 
                   YPGAPGAYPGAPAPGV, 
                   (amino acids 63-78 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                     
                 
                   YPGAPAPGVYPGPPSGPGA, 
                   (amino acids 70-88 of SEQ ID 
                 
                     
                   NO: 1) 
                 
                   and 
                 
                     
                 
                   YPSSGQPSATGA. 
                   (amino acids 89-100 of SEQ 
                 
                     
                   ID NO: 1) 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         4 . The kit of  claim 1  wherein the binding moieties are monoclonal antibodies. 
     
     
         5 . The kit of  claim 4  wherein the monoclonal antibodies are selected from the group consisting of 9H3.2, 87B5, and M3/38. 
     
     
         6 . The kit of  claim 4 , wherein the antibody binds to an epitope comprising MADNFSLHDALSGS (amino acids 1-14 of SEQ ID NO:1), GNPNPQGWPGA (amino acids 15-25 of SEQ ID NO:1), or YPGAPAPGVYPGPPSGPGA (amino acids 70-88 of SEQ ID NO:1). 
     
     
         7 . The kit of  claim 4 , wherein the monoclonal antibody is a mouse antibody. 
     
     
         8 . A method for detecting the level of human galectin-3 in a sample from a subject, the method comprising:
 subjecting the sample to a double binding moiety sandwich assay, at least one of the binding moieties being labeled with a detectable label, each of the binding moieties being specific for respective, non-overlapping epitopes on the N-terminal portion of galectin-3, which comprises the sequence of amino acids:   
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                 
                 
               
                     
                   MADNFSLHDA LSGSGNPNPQ GWPGAWGNQP AGAGGYPGAS 
                 
                     
                     
                 
                     
                   YPGAYPGQAP PGAYPGQAPP GAYPGAPGAY PGAPAPGVYP 
                 
                     
                     
                 
                     
                   GPPSGPGAYP SSGQPSATGA YPATGPYGAP AGP. 
                 
             
                
               
            
             
                
                
                
                
                
               
            
           
         
       
     
     
         9 . The method of  claim 8 , wherein the sandwich assay provides a qualitative result indicative of the presence or absence of a galectin-3 concentration above a threshold. 
     
     
         10 . The method of  claim 9 , wherein the threshold galectin-3 concentration in the sample from the subject is in the range of 15-20 ng/ml. 
     
     
         11 . The method of  claim 9 , wherein the threshold galectin-3 concentration in the sample from the subject is in the range of 20-25 ng/ml. 
     
     
         12 . The method of  claim 9 , wherein the threshold galectin-3 concentration in the sample from the subject is in the range of 25-30 ng/ml. 
     
     
         13 . The method of  claim 9 , wherein the threshold galectin-3 concentration in the sample from the subject is in the range of 30-35 ng/ml. 
     
     
         14 . The method of  claim 8 , wherein the sandwich assay provides a quantitative result indicative of the galectin-3 concentration. 
     
     
         15 . The method of  claim 8  comprising the additional step of comparing the result of the sandwich assay to data correlating the concentration of galectin-3 in the sample with data relating the result with risk of the subject developing symptoms of heart failure (HF) or with severity of diagnosed HF disease of the subject. 
     
     
         16 . The method of  claim 8 , further comprising repeating over time to obtain a trend. 
     
     
         17 . A method for detecting a risk of development or progression of heart failure in a subject, the method comprising determining whether the galectin-3 concentration in a sample from the subject exceeds a threshold of at least 15 ng/ml. 
     
     
         18 . The method of  claim 17 , wherein the threshold is in the range of 15-20 ng/ml. 
     
     
         19 . The method of  claim 17 , wherein the threshold is in the range of 20-25 ng/ml. 
     
     
         20 . The method of  claim 17 , wherein the threshold is in the range of 25-30 ng/ml. 
     
     
         21 . The method of  claim 17 , wherein the threshold is in the range of 30-35 ng/ml.

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