Methods for detecting neutralizing antibodies to parathyroid hormone (pth) and parathyroid hormone-related peptide (pthrp) analog

Abstract

The present disclosure is directed to methods (e.g., in vitro methods) for detecting the presence of neutralizing antibodies to PTH or PTHrP analog in a sample. The in vitro method comprises the steps of obtaining a sample from a subject; contacting the sample with a cell; measuring cyclic adenosine monophosphate (cAMP) levels; and detecting the presence of neutralizing antibodies, when cAMP levels are reduced relative to a negative control sample without neutralizing antibodies. An in vitro method of detecting the presence of neutralizing antibodies in a sample from a subject treated with Abaloparatide is also provided. Further provided herein is a kit for carrying out the methods described herein comprising components required to carry out the obtaining, contacting, measuring and detecting steps and instructions for use.

1 . An in vitro method for detecting the presence of neutralizing antibodies to abaloparatide in a serum sample from a subject treated with abaloparatide, the method comprising:
 obtaining the serum sample from the subject;   preincubating the serum sample with a predetermined amount of abaloparatide for a period of at least 30 minutes;
 contacting the preincubated serum sample with a population of cells or a cell, wherein the cell or cells are rat epithelial cell line UMR-106, and wherein contacting comprises incubating the cell or cells with the serum sample for a period of time; 
   measuring cyclic adenosine monophosphate (cAMP) levels in the serum sample by a competitive immunoassay utilizing an electrochemiluminescent detection method;   detecting the presence of neutralizing antibodies indicated by reduced cAMP levels relative to a negative control sample which does not include neutralizing antibodies; and   discontinuing treating the subject with abaloparatide when neutralizing antibodies are detected in the serum sample.   
     
     
         2 . The method of  claim 1 , wherein the subject is treated with abaloparatide for osteoporosis, the method further comprising monitoring the subject for development of potential immunogenicity. 
     
     
         3 . An in vitro method for detecting the presence of neutralizing antibodies to abaloparatide in a serum sample from a subject treated with abaloparatide, the method comprising:
 obtaining the serum sample from the subject;   preincubating the serum sample with a predetermined amount of abaloparatide for a period of at least 30 minutes;
 contacting the preincubated serum sample with a population of cells or a cell, wherein the cell or cells are rat epithelial cell line UMR-106, and wherein contacting comprises incubating the cell or cells with the serum sample for a period of time; 
   measuring cyclic adenosine monophosphate (cAMP) levels in the serum sample by a competitive immunoassay utilizing an electrochemiluminescent detection method;   detecting the presence of neutralizing antibodies indicated by reduced cAMP levels relative to a negative control sample which does not include neutralizing antibodies; and   increasing or decreasing a dosage of abaloparatide administered to the subject when neutralizing antibodies are detected in the serum sample.   
     
     
         4 . The method of  claim 3 , wherein the subject is treated with abaloparatide for osteoporosis, the method further comprising monitoring the subject for development of potential immunogenicity.