Methods of detecting anti-leptin neutralizing antibodies

Abstract

Provided herein are methods for detecting neutralizing antibodies to leptins, including metreleptin, as well as identifying subjects having such neutralizing antibodies.

1 . A kit for use in detecting neutralizing antibodies to leptin in a sample, comprising (a) labeled leptin, and (b) a substrate capable of binding the labeled leptin. 
     
     
         2 . The kit of  claim 1 , wherein the substrate capable of binding the labeled leptin comprises (i) a coated matrix, and (ii) a tagged leptin receptor bound to the matrix of (i). 
     
     
         3 . The kit of  claim 2 , wherein the substrate capable of binding the labeled leptin comprises (i) a coated matrix, (ii) an antibody bound to the matrix of (i), and (iii) a tagged leptin receptor bound to the antibody of (ii). 
     
     
         4 . The kit of  claim 1 , wherein the labeled leptin is labeled with horseradish peroxidase or ruthenium tris-bipyridine. 
     
     
         5 . The kit of  claim 1 , comprising an acidic solution. 
     
     
         6 . The kit of  claim 1 , wherein said substrate capable of binding the labeled leptin comprises:
 (i) a coated matrix; and   (ii) a tagged leptin receptor bound to the matrix of (i).   
     
     
         7 . The kit of  claim 6 , wherein the coated matrix is a streptavidin-coated matrix. 
     
     
         8 . The kit of  claim 2 , wherein the leptin receptor is tagged with biotin. 
     
     
         9 . The kit of  claim 1 , wherein said substrate capable of binding the labeled leptin comprises:
 (i) a coated matrix;   (ii) an antibody bound to the matrix of (i); and   (iii) a tagged leptin receptor bound to the antibody of (ii).   
     
     
         10 . The kit of  claim 9 , wherein the coated matrix is a streptavidin coated matrix. 
     
     
         11 . The kit of  claim 9 , wherein the tagged leptin receptor comprises a tag suitable for affinity purification. 
     
     
         12 . The kit of  claim 11 , wherein the tagged leptin receptor comprises a hexa-histidine tag. 
     
     
         13 . The kit of  claim 9 , wherein the antibody is labeled with a tag suitable to bind to the coated matrix. 
     
     
         14 . The kit of  claim 13 , wherein the tag is biotin. 
     
     
         15 . The kit of  claim 1 , wherein the labeled leptin comprises recombinant human leptin. 
     
     
         16 . The kit of  claim 1 , wherein the labeled leptin comprises metreleptin. 
     
     
         17 . The kit of  claim 5 , wherein the acidic solution is an acetic acid solution. 
     
     
         18 . The kit of  claim 17 , wherein the acidic solution is 0.8% acetic acid.