人甲状腺过氧化物酶的提取、纯化及鉴定

EXTRACTION AND PURIFICATION OF HUMAN THYROID PEROXIDASE

目的：提取较纯的ｈＴＰＯ，为甲状腺激素合成过程的研究和建立ｈＴＰＯＡｂ检测方法提供了实验材料。方法：本文采用胰蛋白酶和脱氧胆酸钠处理手术切除人甲状腺组织，经超速离心和ＳｅｐｈａｃｒｙｌＳ－３００柱层析提纯人甲状腺过氧化物酶（ｈＴＰＯ），并进行了愈创木酚氧化法ｈＴＰＯ酶活性测定。结果：本研究获得的ｈＴＰＯ比生物活性为８．４３愈创木酚单位（ＧＵ）／ｍｇ。经ＳＤＳ－ＰＡＧＥ电泳鉴定示提纯的ｈＴＰＯ在分子量１０７ｋＤ处有一清晰色带，而于９３ｋＤ处也有一色带，但较浅。结论：该方法提纯ｈＴＰＯ操作简便、快速，获得的ｈＴＰＯ较纯。

Objective:It aimed at obfaining hTPOThe hTPO would provied the experimental material for studies of thyroid hormone syntehesis and establishment of antihTPO antibody measurementMethods:After human thyroid tissue removed in operation was treated by trypsin and deoxycholic acid sodium,human thyroid peroxidase(hTPO) were extracted by ultracetrifugation and purified by chromatography of Sephacryl S300 columnEnzyme activity of hTPO was measured by guaiacal oxidationResults:Results showed bioactivity of the hTPO was 843 Guaiacal Unit (GU)/mg It was found that there was a band in molecule weight 107kD,and there was also a band in molecule weight 93kD which was lightConclusions:This method of the hTPO extraction and purification was easy and quick to operat