Lissens W, Zenati A, Liebaers I
Biochim Biophys Acta. 1984 Oct 16;801(3):365-71. doi: 10.1016/0304-4165(84)90140-5.
Human iduronate 2-sulphate sulphatase (EC 3.1.6.-) from urine has been purified by affinity chromatography on concanavalin A-Sepharose, ammonium sulphate fractionation and DEAE-cellulose chromatography. With ion-exchange chromatography, the enzyme was resolved in two activity peaks. The less anionic of these forms was further purified by polyacrylamide gel electrophoresis under non-denaturing conditions. Anti-iduronate 2-sulphate sulphatase antibodies were obtained from mice immunized with polyacrylamide eluted enzyme. The specificity of the antibodies towards iduronate 2-sulphate sulphatase was demonstrated by immunoprecipitation of the enzyme from partially purified urine protein. The procedure described in this work opens the way to the application of hybridoma technology to iduronate 2-sulphate sulphatase.
已通过伴刀豆球蛋白A-琼脂糖亲和层析、硫酸铵分级分离和DEAE-纤维素层析对人尿中的艾杜糖醛酸2-硫酸酯酶(EC 3.1.6.-)进行了纯化。通过离子交换层析,该酶被分离为两个活性峰。这些形式中阴离子性较弱的一种通过非变性条件下的聚丙烯酰胺凝胶电泳进一步纯化。用聚丙烯酰胺洗脱的酶免疫小鼠,获得了抗艾杜糖醛酸2-硫酸酯酶抗体。通过从部分纯化的尿蛋白中免疫沉淀该酶,证明了抗体对艾杜糖醛酸2-硫酸酯酶的特异性。本研究中描述的方法为将杂交瘤技术应用于艾杜糖醛酸2-硫酸酯酶开辟了道路。