Freeze H H
La Jolla Cancer Research Foundation, La Jolla, California, USA.
Curr Protoc Mol Biol. 2001 May;Chapter 17:Unit17.1. doi: 10.1002/0471142727.mb1701s22.
This unit begins by describing some properties of glycoproteins (e.g., subcellular location and solubility) that may be useful in determining which purification techniques to try. This discussion is followed by two protocols describing preparative glycoprotein purification using lectin-affinity chromatography, as well as an outline for a small-scale pilot procedure designed to check lectin binding and elution conditions. Lectins are often used for purifying glycoproteins because, in contrast to conventional purification procedures (e.g., gel filtration and ion-exchange chromatography) that exploit general physical properties of glycoproteins, lectins recognize specific three-dimensional structures created by a cluster of sugar residues. Conventional purification procedures are generally tried before applying lectin-affinity chromatography.
本单元首先描述了糖蛋白的一些特性(如亚细胞定位和溶解性),这些特性可能有助于确定尝试使用哪种纯化技术。接下来是两个使用凝集素亲和色谱法进行制备性糖蛋白纯化的实验方案,以及一个用于检查凝集素结合和洗脱条件的小规模预实验程序概述。凝集素常用于纯化糖蛋白,因为与利用糖蛋白一般物理性质的传统纯化程序(如凝胶过滤和离子交换色谱法)不同,凝集素能识别由一簇糖残基形成的特定三维结构。在应用凝集素亲和色谱法之前,通常先尝试传统纯化程序。