Induction of tsh binding inhibitory immunoglobulins with the extracellular domain of human thyrotropin receptor produced using baculovirus expression system

G. S. Seetharamaiah, Rajesh K. Desai, John S. Dallas, Kazuo Tahara, Leonard D. Kohn, Bellur S. Prabhakar

Research output: Contribution to journalArticle

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Abstract

A cDNA encoding the extracellular domain of human TSHr (ETSHr) was expressed in large quantities using the baculovirus expression system. Maximum level of protein was produced at 60 hr post-infection and represented approximately 20% of the total cellular protein. The identity of the protein as ETSHr was confirmed by Western blot using antibodies to synthetic peptides derived from the TSHr. The protein has an apparent molecular weight of 50 kDa and is larger than the predicted size of 44 ma, suggesting that the protein is glycosylated. Polyclonal antibodies raised in rabbits against gel purified ETSHr blocked the binding of 125I-TSH to native TSHr in solubilized porcine thyroid membranes in a radioreceptor assay. The availability of this antigenically active protein will facilitate further characterization of the protein and analysis of immune response against TSHr in experimental animals as well as in patients with autoimmune Graves' disease.

Original languageEnglish (US)
Pages (from-to)315-320
Number of pages6
JournalAutoimmunity
Volume14
Issue number4
DOIs
StatePublished - Jan 1 1993

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Keywords

  • Baculovirus expression
  • Graves' disease
  • TSH binding inhibitory immunoglobulins
  • Thyrotropin receptor

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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