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Solubilization of Human Thyroid Microsomal Antigen^*

Cattedra di Patologia Speciale Medica II, University of Pisa Pisa, Italy

Address all correspondence and requests for reprints to: Dr. Aldo Pinchera, Patologia Speciale Medica II, Via Bonanno 48, 56100, Pisa, Italy.

The ability of detergents (Triton X-100 and deoxycholate), high ionic strength solution (3 M KC1), 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 ^l25I-labeled human immunoglobulin G (IgG) with elevated antimicrosomal (anti-M) and undetectable antithyroglobulin antibodies (anti-M IgG). All solubilized materials were shown to bind [^l25I]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 KC1 in decreasing order. Affinity chromatography with the deoxycholate-solubilized material coupled to Sepharose 4B resulted in a 15.6-fold purification of [^l25I]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. (J Clin Endocrinol Metab 48: 207, 1979)

^* This work was supported by grants from Consiglio Nazionale delle Ricerche, Rome, Italy.

Received May 14, 1978.