Solubilization of human thyroid microsomal antigen

S. Mariotti, A. Pinchera, C. Marcocci, P. Vitti, C. Urbano, L. Chiovato, M. Tosi, L. Baschieri

Research output: Contribution to journalArticle

Abstract

The ability of detergents (Triton X-100 and deoxycholate), high ionic strength solution (3 M KCl), and proteolytic enzymes (papain and trypsin) to solubilize human thyroid microsomal antigen was studied. Antigenic activity released from thyroid microsomal preparation into the incubation mixture was separated by centrifugation at 143,000 x g for 90 min and measured using 125I-labeled human immunoglobulin G (IgG) with elevated antimicrosomal (anti-M) and undetectable antithyroglobulin antibodies (anti-M IgG). All solubilized materials were shown to bind [ 125I]anti-M IgG and to inhibit its binding to untreated thyroid microsomes. These effects were specific and dose related. Measurements of specific activity and total amount of solubilized antigen by an absorption technique showed that Triton X-100 was the most effective agent, followed by deoxycholate, papain, trypsin, and 3 M KCl in decreasing order. Affinity chromatography with the deoxycholate-solubilized material coupled to Sepharose 4B resulted in a 15.6-fold purification of [ 125I]anti-M antibodies. The present results indicate that thyroid microsomal antigen may be solubilized by several agents and this can provide the basis for its identification and purification.

Original languageEnglish
Pages (from-to)207-212
Number of pages6
JournalJournal of Clinical Endocrinology and Metabolism
Volume48
Issue number2
Publication statusPublished - 1979

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

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    Mariotti, S., Pinchera, A., Marcocci, C., Vitti, P., Urbano, C., Chiovato, L., Tosi, M., & Baschieri, L. (1979). Solubilization of human thyroid microsomal antigen. Journal of Clinical Endocrinology and Metabolism, 48(2), 207-212.