Novel thyroid peroxidase autoantibody immunoassay

Abstract

Disclosed herein are immunoassays for detecting an anti-thyroid peroxidase antibody in a biological sample from a subject and/or diagnosing a thyroid disease in a subject. The disclosed immunoassays employ a recombinant cynomolgus monkey thyroid peroxidase (rTPO) and assess the level of anti-thyroid peroxidase antibody-induced formation or disruption of complexes comprising a solid support and the rTPO.

What is claimed is: 
     
         1 . A method of diagnosing a thyroid disease in a subject, the method comprising:
 a) incubating a biological sample from the subject with:
 a solid support, 
 an unlabeled recombinant cynomolgus monkey thyroid peroxidase (rTPO), and 
 a labeled cynomolgus monkey rTPO, 
 wherein, in the presence of an anti-thyroid peroxidase antibody in the biological sample, a complex comprising the labeled cynomolgus monkey rTPO, the unlabeled cynomolgus monkey rTPO, and the solid support is formed; and 
   b) diagnosing the subject with the thyroid disease if the complex is formed.   
     
     
         2 . A method of diagnosing a thyroid disease in a subject, the method comprising:
 a) incubating a biological sample from the subject with:
 a solid support, 
 a cynomolgus monkey rTPO, and 
 an anti-human secondary antibody, 
 wherein, in the presence of an anti-thyroid peroxidase antibody in the biological sample, a complex comprising the solid support, the cynomolgus monkey rTPO, and the anti-human secondary antibody is formed; and 
   b) diagnosing the subject with the thyroid disease if the complex is formed.   
     
     
         3 . A recombinantly produced thyroid peroxidase (rTPO) comprising the amino acid sequence of SEQ ID NO: 1. 
     
     
         4 . The rTPO of  claim 3 , wherein the rTPO comprises a label. 
     
     
         5 . The rTPO of  claim 4 , wherein the label comprises an enzyme conjugate, a fluorescent probe, a radioactive isotope, a chemiluminescent compound, a bioluminescent compound, or a combination thereof. 
     
     
         6 . The rTPO of  claim 5 , wherein the label comprises an acridinium ester (AE) or an analog thereof. 
     
     
         7 . The rTPO of  claim 6 , wherein the acridinium ester analog is DMAE, NSP-DMAE, HQYAE, ZAE, Iso-Di-ZAE, was TSP-AE, or HEG-GLU-AE. 
     
     
         8 . The rTPO of  claim 4  in complex with an anti-thyroid peroxidase antibody. 
     
     
         9 . A kit comprising:
 a solid support;   a cynomolgus monkey rTPO; and   an anti-human secondary antibody.   
     
     
         10 . The kit of  claim 9 , wherein the solid support comprises a column matrix material, a culture plate, a tube, a dish, a flask, a microtiter plate, a bead, or a combination thereof. 
     
     
         11 . The kit of  claim 9 , wherein:
 the cynomolgus monkey rTPO comprises a label; or   the anti-human secondary antibody comprises a label.   
     
     
         12 . The kit of  claim 11 , wherein the label comprises an enzyme conjugate, a fluorescent probe, a radioactive isotope, a chemiluminescent compound, a bioluminescent compound, or a combination thereof. 
     
     
         13 . The kit of  claim 12 , wherein the label is an acridinium ester (AE) or an analog thereof. 
     
     
         14 . A kit comprising:
 a solid support;   a cynomolgus monkey rTPO; and   an anti-TPO antibody.   
     
     
         15 . The kit of  claim 14 , wherein the solid support comprises a column matrix material, a culture plate, a tube, a dish, a flask, a microtiter plate, a bead, or a combination thereof. 
     
     
         16 . The kit of  claim 14 , wherein a portion of the anti-TPO antibodies are unlabeled and a portion of the anti-TPO antibodies are labeled. 
     
     
         17 . The kit of  claim 16 , wherein the label comprises an enzyme conjugate, a fluorescent probe, a radioactive isotope, a chemiluminescent compound, a bioluminescent compound, or a combination thereof. 
     
     
         18 . The kit of  claim 17 , wherein the label is an acridinium ester (AE) or an analog thereof. 
     
     
         19 . A kit comprising:
 a solid support,   an unlabeled anti-TPO antibody,   a cynomolgus monkey rTPO,   a labeled anti-TPO antibody, and   an ADVIA Centaur© or Atellica IM© ReadyPack©.