Parker N, Finnie I A, Raouf A H, Ryder S D, Campbell B J, Tsai H H, Iddon D, Milton J D, Rhodes J M
University Department of Medicine, Royal Liverpool University Hospital, UK.
Biomed Chromatogr. 1993 Mar-Apr;7(2):68-74. doi: 10.1002/bmc.1130070204.
High performance gel filtration on monodisperse cross-linked agarose (Superose 6) has been assessed as a system for purification of mucus glycoproteins. Comparison with the conventional two-step purification of mucus glycoprotein by Sepharose CL4B gel filtration followed by caesium chloride density gradient centrifugation shows that purification by high performance gel filtration is at least as thorough, yielding mucin that is free from non-mucin glycoproteins as defined by buoyant density, mobility on sodium dodecyl sulphate-polyacrylamide gel electrophoresis and absence of Concanavalin A binding (mannose-containing) material. This technique allows mucus glycoprotein to be purified from lyophilized crude mucin in 120 min compared with approximately 72 h using the conventional techniques. This makes the comparative study of mucus glycoprotein changes in disease states much more feasible.
已评估了在单分散交联琼脂糖（Superose 6）上进行的高效凝胶过滤作为一种纯化黏液糖蛋白的系统。将其与通过Sepharose CL4B凝胶过滤随后进行氯化铯密度梯度离心的传统两步法纯化黏液糖蛋白进行比较，结果表明，通过高效凝胶过滤进行的纯化至少同样彻底，所产生的黏蛋白不含非黏蛋白糖蛋白，这是根据浮力密度、在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上的迁移率以及缺乏伴刀豆球蛋白A结合（含甘露糖）物质来定义的。与使用传统技术大约72小时相比，该技术可在120分钟内从冻干的粗黏蛋白中纯化出黏液糖蛋白。这使得对疾病状态下黏液糖蛋白变化的比较研究更加可行。
