Biologically active synthetic thyrotropin and cloned gene for producing same

Abstract

Substantially pure recombinant TSH has been prepared from a clone comprising complete nucleotide sequence for the expression of the TSH. Diagnostic and therapeutic applications of the synthetic TSH are described.

1 . Substantially pure, biologically active recombinant human thyrotropin (rTSH).  
   
   
       2 . The thyrotropin of  claim 1  labeled isotopically with  131 I,  125 I, chemiluminiscently or fluorescently.  
   
   
       3 . The thyrotropin of  claim 1  produced by recombinant genetic process using constructs with gene elements that enhance thyrotropin production.  
   
   
       4 . A clone comprising complete nucleotide sequence for the expression of the thyrotropin of  claim 1  in a suitable expression vector.  
   
   
       5 . The clone of  claim 4  further comprising first untranslated exon of TSH-β.  
   
   
       6 . A method for producing TSH, comprising: 
 (a) allowing expression of TSH by the clone of  claim 4  in a suitable expression vector; and    (b) then recovering substantially pure TSH by conventional purification and isolation methodology.    
   
   
       7 . A method for producing TSH, comprising: 
 (a) allowing expression of TSH by the clone of  claim 5  in a suitable expression vector; and    (b) then recovering substantially pure TSH by conventional purification and isolation methodology.    
   
   
       8 . A method for producing TSH, comprising: 
 (a) allowing expression of TSH by the clone of  claim 4 , wherein TSHα is about 3 to 5 times in excess of TSHβ;    (b) then recovering substantially pure TSH by conventional purification and isolation methodology.    
   
   
       9 . A method for producing TSH, comprising: 
 (a) allowing expression of TSH by the clone of  claim 5 , wherein TSHα is about 3 to 5 times in excess of TSHβ;    (b) then recovering substantially pure TSH by conventional purification and isolation methodology.    
   
   
       10 . A TSH antagonist produced by a mutant of the clone of  claim 4 .  
   
   
       11 . A TSH agonist produced by a mutant of the clone of  claim 4 .  
   
   
       12 . A kit comprising containers separately containing: 
 (a) universal standard of substantially pure unlabeled rTSH;    (b) substantially pure, labeled rTSH;    (c) antibodies against purified rTSH; and    (d) instructional material describing the use of reagents (a), (b) and (c).    
   
   
       13 . Anti-rTSH antibodies without interfering cross-reactivity with non-TSH hormones.  
   
   
       14 . A method for determining the level of TSH in a sample, comprising reacting an aliquot of a sample in which the amount of TSH is to be determined with the antibodies of  claim 13  and comparing the level of antibody reactivity with a predetermined standard antibody-rTSH reactivity curve to determine the amount of TSH present in said sample.  
   
   
       15 . A method of diagnosing the extent of thyroid cancer, comprising administering rTSH of  claim 1  to a patient to maximize  131 I uptake, and then administering a visualizing dose of  131 I to said patient; and then visualizing the cancer by standard visualizing means.  
   
   
       16 . A method for treating thyroid cancer, comprising administering therapeutic regimen of a combination of rTSH of  claim 1  and  131 I or  131 I-labeled rTSH to a patient afflicted with thyroid cancer.  
   
   
       17 . A method of blocking TSH activity, comprising inhibiting TSH activity by competitive amount of the antagonist of  claim 10 .  
   
   
       18 . A method of stimulating TSH activity, comprising inducing TSH production by the agonist of  claim 11.