Procedure for isolation of gangliosides in high yield and purity: simultaneous isolation of neutral glycosphingolipids.
Procedure for isolation of gangliosides in high yield and purity: simultaneous isolation of neutral glycosphingolipids.
复制标题
高产率和高纯度分离神经节苷脂的程序：同时分离中性糖鞘脂。
DOI：
10.1016/0003-2697(85)90641-4
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发表时间：
1985
影响因子：
2.9
通讯作者：
Ledeen,RW
中科院分区：
文献类型：
--
作者：
Byrne,MC;Sbaschnig-Agler,M;Aquino,DA;Sclafani,JR;Ledeen,RW
虽然已经描述了用于神经节苷脂提取和分离的几种方法，但相对较少关注从肽、磷脂和各种低分子量污染物中分离的有效性。描述了一种从1- 400-mg样品（湿重）中分离高纯度和良好收率的神经节苷脂的方法。一个关键的步骤是温和的酸化后均质化，旨在解离神经节苷脂从亲脂性肽共提取到有机溶剂。这已被证明对髓磷脂和含髓磷脂的组织（例如，白色物质、神经），其蛋白质在神经节苷脂分离中存在特殊问题。在这项研究中，分离的影响，通过连续的色谱Sephadex LH-20，DEAE-Sephadex，和硅胶后的初始酸化。应用于牛白色物质的方法得到的组织浓度（根据产率和放射性标记示踪剂回收率计算）与采用三种先前描述的程序获得的浓度相似;然而，肽污染物的数量级较低。去除低分子量的污染物，包括核苷酸糖，几乎是完全的。除神经节苷脂分离外，该方法还可用于获得中性鞘糖脂。它被认为对多种组织具有广泛的适用性。
While several methods for ganglioside extraction and isolation have been described, relatively little attention has been given to the effectiveness of separation from peptides, phospholipids, and various low-molecular-weight contaminants. A procedure is described for isolation of gangliosides in high purity and good yield from 1- to 400-mg samples (wet wt). A key step was mild acidification following homogenization, designed to dissociate gangliosides from lipophilic peptides which coextracted into organic solvents. This has proved particularly helpful for myelin and myelin-containing tissues (e.g., white matter, nerve) whose proteins have presented special problems in ganglioside isolation. In this study isolation was effected by consecutive chromatographies on Sephadex LH-20, DEAE-Sephadex, and silica gel following the initial acidification. The method applied to bovine white matter gave tissue concentrations (calculated from yields and radiolabeled tracer recoveries) that were similar to those obtained with three previously described procedures; however, peptide contaminants were an order of magnitude lower. Removal of low-molecular-weight contaminants, including nucleotide sugars, was virtually complete. In addition to ganglioside isolation the method can be used to obtain neutral glycosphingolipids as well. It is believed to have broad applicability to a diversity of tissues.