Properties of thyroglobulins from normal thyroid and thyroid tumor on a concanavalin A-sepharose column.
Affinity chromatography on a concanavalin A (con A)-Sepharose column is a potentially useful for the isolation of whole thyroglobulin (Tg) at least from normal thyroid tissue. In addition to being a simple procedure for the isolation of Tg, large amounts of Tg can be applied to the column and recovered in good yield with a buffer containing MeG. In gradient elution with buffer containing increasing amounts of MeG, a single but broad peak was obtained, without separation into subfractions. However, a hemagglutination-inhibition test showed that the Tg preparation eluted early from the column had less affinity for con A than the Tg preparation eluted later, suggesting a heterogeneous distribution of carbohydrate moieties among Tg preparations. When human Tg from thyroid tumor was applied to the column, tumor Tg partly passed through the column without being adsorbed. This unadsorbed Tg showed a very low affinity for lectins, con A and wheat germ agglutinin (WGA), as determined by a double diffusion reaction in agar gel. In contrast to this fraction, the Tg adsorbed on the con A-gel column showed a very strong affinity for WGA, differing from Tg of normal thyroid tissue. Therefore, tumor Tg preparation appears to have an abnormally modified carbohydrate structure, at least in part. The higher affinity for WGA (with a specificity for N-acetylglucosamine) seen in adsorbed Tg could be due to a larger amount of GlcNAc residues which bind irregularly in the carbohydrate moiety of tumor Tg.
D000236:Adenoma / Q000032:analysis; D000818:Animals; D002277:Carcinoma / Q000032:analysis; D002846:Chromatography, Affinity; D006801:Humans; D005779:Immunodiffusion; D008297:Male; D009919:Orchiectomy; D012016:Reference Values; D012685:Sepharose / Q000031:analogs & derivatives; D013552:Swine; D013954:Thyroglobulin / Q000302:isolation & purification*; D013961:Thyroid Gland / Q000032:analysis*; D013964:Thyroid Neoplasms / Q000032:analysis*
C028326:concanavalin A-sepharose; D013954:Thyroglobulin; D012685:Sepharose