Generation of a mouse monoclonal TSH receptor antibody with stimulating activity

S. Costagliola, J. D F Franssen, M. Bonomi, E. Urizar, M. Willnich, A. Bergmann, G. Vassart

Research output: Contribution to journalArticlepeer-review

Abstract

A Balb/c mouse was subjected to genetic immunization with a cDNA construct encoding the human thyrotropin receptor (TSHr). The immune response of the mouse resulted in the production of immunoglobulins recognizing the TSHr in three different assays: (1) flow immunocytometry (FACS) with CHO cells expressing the receptor; (2) receptor-dependent stimulation of cAMP production in the same cell line; and (3) competition with labeled TSH for binding to the receptor. One thousand hybridomas were generated from the spleen of the mouse and their supernatants were screened. A single monoclonal, IRI-SAb1, scored positive in all three assays and was studied further. It stimulated 13-fold cAMP production in TSHr-expressing CHO cells, with an EC50 in the low nanomolar range. When compared with bovine TSH, IRI-SAb1 behaved as a partial agonist. Contrary to the expectation from the characteristic of autoantibodies of Graves' patients, IRI-SAb1 recognized a linear epitope, which was localized in a segment encompassing the first 281 residues of the receptor.

Original languageEnglish
Pages (from-to)891-896
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume299
Issue number5
DOIs
Publication statusPublished - 2002

Keywords

  • Graves' autoantibodies
  • Monoclonal
  • TSAb
  • TSH receptor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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