Recombinant thyrotropin containing a β-subunit chimera with the human chorionic gonadotropin-β carboxy-terminus is biologically active, with a prolonged plasma half-life: Role of carbohydrate in bioactivity and metabolic clearance

Lata Joshi, Yoko Murata, Fredric E. Wondisford, Mariusz W. Szkudlinski, Rajesh Desai, Bruce D. Weintraub

Research output: Contribution to journalArticle

Abstract

Recombinant TSH is now successfully being used in clinical studies of thyroid cancer. Because of its therapeutic potential, we have constructed a longer acting analog of TSH by fusing the carboxy-terminal extension peptide (CTEP) of hCG β onto TSH β. When coexpressed either withα-subunit complementary DNA orα minigene in African green monkey (COS-7) and human embryonic kidney (293) cells, the chimera was fully bioactive in vitro and exhibited enhanced in vivo potency associated with a prolonged plasma half-life. The addition of 25 amino acids with 4 O-linked oligosaccharide chains did not affect the assembly and secretion of chimeric TSH. Wild-type (WT) and chimeric TSH secreted by COS-7 and 293 cells displayed wide differences in their plasma half-lives, presumably due to the presence of terminal sialic acid and SO4 on their oligosaccharide chains, respectively. Chimeric and WT TSH secreted by both cell lines demonstrated similar bioactivity in cAMP production, with some differences in [3H]thymidine incorporation. Chimeric TSH appears to be more effective in COS-7 cells than in 293 cells, as judged by growth assay. COS-7-produced chimeric TSH showed the maximum increase in half-life, indicating the importance of sialic acid in prolonging half-life and in vivo potency. Sulfation of both subunits, predominantly β and to a lesser extentα, appears to be responsible at least in part for the increased metabolic clearance of WT and chimeric TSH secreted by 293 cells. Apart from its therapeutic potential, chimeric TSH produced in various cell lines can be used as a tool to delineate the roles of sulfate and sialic acid in the in vivo clearance and, thereby, the in vivo bioactivity.

Original languageEnglish (US)
Pages (from-to)3839-3848
Number of pages10
JournalEndocrinology
Volume136
Issue number9
DOIs
StatePublished - Sep 1995

    Fingerprint

ASJC Scopus subject areas

  • Endocrinology

Cite this