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.

What is claimed is:  
     
         1 . Substantially pure, biologically active recombinant human thyrotropin (rTSH).  
     
     
         2 . The thyrotropin of    claim 1    labeled isotopically with I 131 , I 125 , chemoluminiscently or fluoroscently.  
     
     
         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   .