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US 6,284,491 B1Granted
Biologically active synthetic thyrotropin and cloned gene for producing same
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorsFredric E Wondisford, Bruce D. Weintraub
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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.
Claims4
A method for producing recombinant human TSH comprising the steps of:
- a) transfecting a eukaryotic cell with an expression vector, wherein said expression vector comprises a contiguous genomic fragment of the human thyrotropin (TSH) β-subunit gene wherein the fragment comprises both coding exons of the TSH β-subunit gene separated by the intron;
- b) culturing said transfected cell under conditions that allow said vector to direct the machinery endogenous in said eukaryotic cell to perform the splicing necessary for expression of mRNA to bring about the production of TSH β-subunit and that allow for the production of TSH α subunit for combination therewith;
- and
- c) recovering substantially pure recombinant human TSH.
1 more element
A method for producing recombinant human TSH comprising the steps of:
- a) transfecting a eukaryotic cell with an expression vector, wherein said expression vector comprises a contiguous genomic fragment of the human thyrotropin (TSH) β-subunit gene wherein the fragment comprises both coding exons of the TSH β-subunit gene separated by the intron and further comprises the 5′ untranslated exon;
- b) culturing said transfected cell under conditions that allow said vector to direct the machinery endogenous in said eukaryotic cell to perform the splicing necessary for expression of mRNA to bring about the production of TSH β-subunit and that allow for the production of TSH α subunit for combination therewith, and
- c) recovering substantially pure recombinant human TSH.
Dependencies are inferred from the claim text.