Serafini-Cessi F, Bellabarba G, Malagolini N, Dall'Olio F
Dipartimento di Patologia Sperimentale, Università di Bologna, Italy.
J Immunol Methods. 1989 Jun 21;120(2):185-9. doi: 10.1016/0022-1759(89)90241-x.
An isolation method for Tamm-Horsfall protein is described which is based on the observation that a diatomaceous earth filter is able to retain most of the glycoprotein present in urine and that the glycoprotein is easily desorbed from the filter by deionized water. This behaviour depends on the tendency of Tamm-Horsfall glycoprotein at normal urinary concentrations to form a gel in a solution containing mono- and divalent ions. By means of two-step filtration, the glycoprotein was purified to homogeneity. The yield was of about 20 mg/l of urine, and the time required for the isolation was approximately 5-6 h. This procedure should be particularly useful for preparing large amounts of Tamm-Horsfall glycoprotein oligosaccharides in order to investigate their potential use as immunosuppressive agents both in vitro and in vivo.
本文描述了一种分离Tamm-Horsfall蛋白的方法,该方法基于以下观察结果:硅藻土过滤器能够保留尿液中存在的大部分糖蛋白,并且该糖蛋白很容易被去离子水从过滤器上解吸下来。这种行为取决于正常尿液浓度下Tamm-Horsfall糖蛋白在含有单价和二价离子的溶液中形成凝胶的倾向。通过两步过滤,糖蛋白被纯化至同质。产量约为每升尿液20毫克,分离所需时间约为5-6小时。该方法对于制备大量Tamm-Horsfall糖蛋白寡糖特别有用,以便研究它们在体外和体内作为免疫抑制剂的潜在用途。