Sethi Manveen K, Fanayan Susan
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.
Australian School of Advanced Medicine, Macquarie University, Sydney, Australia.
Curr Protoc Protein Sci. 2015 Aug 3;81:24.6.1-24.6.10. doi: 10.1002/0471140864.ps2406s81.
Lectins are sugar-binding proteins that can recognize and bind to carbohydrates conjugated to proteins and lipids. Coupled with mass spectrometry technologies, lectin affinity chromatography is becoming a popular approach for identification and quantification of glycoproteins in complex samples such as blood, tumor tissues, and cell lines. Given the commercial availability of a large number of lectins that recognize diverse sugar structures, it is now possible to isolate and study glycoproteins for biological and medical research. This unit provides a general guide to single-lectin-based enrichment of glycoproteins from serum-free conditioned media. Due to the unique carbohydrate specificity of most lectins and the complexity of the samples, optimization steps may be required to evaluate different elution buffers and methods as well as binding conditions, for each lectin, for optimal recovery of bound glycoproteins.
凝集素是一类能识别并结合与蛋白质和脂质结合的碳水化合物的糖结合蛋白。结合质谱技术，凝集素亲和色谱法正成为一种用于鉴定和定量分析血液、肿瘤组织及细胞系等复杂样品中糖蛋白的常用方法。鉴于市面上有大量能识别多种糖结构的凝集素可供选择，目前已能够分离并研究用于生物学和医学研究的糖蛋白。本单元提供了一个从无血清条件培养基中基于单一凝集素富集糖蛋白的通用指南。由于大多数凝集素独特的碳水化合物特异性以及样品的复杂性，可能需要优化步骤来评估不同的洗脱缓冲液、方法以及每种凝集素的结合条件，以实现结合糖蛋白的最佳回收率。
