Paterson A J, Hearn M T
Aust J Exp Biol Med Sci. 1979 Dec;57(6):641-9. doi: 10.1038/icb.1979.68.
The application of fused rocket immunoelectrophoretic methods for the analysis of human and sheep thyroid extracts is reported. Different experimental procedures, evaluating thyroglobulin preparations obtained by gel chromatographic separations with a variety of gel supports and elution conditions, were examined. Adsorption experiments demonstrated that many of the antigenic contaminants in chromatographed thyroglobulin were due to serum proteins. The precipitin profiles obtained with the fused rocket immunoelectrophoretic experiments indicate that considerable tailing of 19S-thyroglobulin occurs when chromatographed on either Sephadex G-200 or Bio-Gel A-1.5 m using low ionic strength buffers. The retention of thyroglobulin on these supports would account, in part, for the observed presence of thyroid proteins with antigenic determinants identical to 19S-thyroglobulin in chromatographed subfractions of notionally lower molecular weight. Because of the case of execution, the described methods provide a useful alternative to existing methods for the assessment of homogeneity of chromatographed thyroglobulin samples.
本文报道了融合火箭免疫电泳法在分析人及羊甲状腺提取物中的应用。研究了不同的实验程序，评估通过凝胶色谱分离法，在多种凝胶支持物及洗脱条件下获得的甲状腺球蛋白制剂。吸附实验表明，经色谱分离的甲状腺球蛋白中的许多抗原性污染物源自血清蛋白。融合火箭免疫电泳实验所得的沉淀素图谱表明，当使用低离子强度缓冲液在Sephadex G - 200或Bio - Gel A - 1.5m上进行色谱分离时，19S - 甲状腺球蛋白会出现相当程度的拖尾现象。甲状腺球蛋白在这些支持物上的保留，部分解释了在理论上分子量较低的色谱亚组分中观察到的具有与19S - 甲状腺球蛋白相同抗原决定簇的甲状腺蛋白的存在。由于操作简便，所述方法为评估经色谱分离的甲状腺球蛋白样品的同质性提供了一种有用的替代现有方法。
