Expression of recombinant human thyrotropin receptor in myeloma cells

Takao Matsuba, Masayuki Yamada, Hideo Suzuki, Akira Kanai, Osamu Isozaki, Tadashi Yoshida, Toshio Tsushima, Kiyoshi Yasukawa

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18 Citations (Scopus)

Abstract

Starting with a previously isolated cDNA for human thyrotropin receptor (TSHR), we established a transformed myeloma cell line, SP56, which expresses human TSHR on its cell surface. Binding analysis showed that SP56 bears 1.1×106TSHR per cell with a Kd of 2.2×10-10M. Using the purified cellular membrane, we established a TSH binding inhibition immunoglobulin (TBII) assay for autoantibodies against TSHR. We compared it with the TBII assay utilizing porcine thyroid membranes expressing porcine TSHR, which has been widely used for TBII assay, by using 96 serum samples from patients with autoimmune thyroid disease and normal individuals. Our TBII assay was more sensitive than the one using porcine TSHR: of 38 sera of patients which were judged negative for autoantibodies to TSHR(TBII value below 10%) by the latter assay, 28 were positive (above 20%) in our assay. By using a perfusion culture system, we obtained as many as 3×1010SP56 cells, from which 3, 450 mg protein of the membrane could be purified; this is sufficient for 15,000 assays. The results indicate that the membrane of the myeloma cell line SP56 is more suitable for use in the TBII assay than the porcine thyroid membrane, in terms of sensitivity to autoantibodies against TSHR in human sera.

Original languageEnglish
Pages (from-to)265-270
Number of pages6
JournalJournal of biochemistry
Volume118
Issue number2
DOIs
Publication statusPublished - 1995 Aug

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Keywords

  • Autoantibody
  • CHO cell, Graves' disease
  • Key words:
  • Myeloma cell
  • Thyrotropin receptor

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Matsuba, T., Yamada, M., Suzuki, H., Kanai, A., Isozaki, O., Yoshida, T., Tsushima, T., & Yasukawa, K. (1995). Expression of recombinant human thyrotropin receptor in myeloma cells. Journal of biochemistry, 118(2), 265-270. https://doi.org/10.1093/oxfordjournals.jbchem.a124901