Willats William G T, Rasmussen Svend Erik, Kristensen Tina, Mikkelsen Jørn Dalgaard, Knox J Paul
Centre for Plant Sciences, University of Leeds, U.K.
Proteomics. 2002 Dec;2(12):1666-71. doi: 10.1002/1615-9861(200212)2:12<1666::AID-PROT1666>3.0.CO;2-E.
The development of DNA and protein microarrays represents a significant advance in transcriptomics and proteomics research. Such arrays allow the high-throughput, parallel analysis of protein occurrence and interactions and gene expression. However, this advance has not been matched by equivalent technology for analysis of glycomes. One reason for this is that compared to proteins, it is difficult to reliably immobilise populations of chemically and structurally diverse glycans. We describe the development of a new microarray slide surface to which diverse glycan structures can be directly immobilised without prior derivatisation of the slide surface or any modification of the arrayed samples. The slides can be used to produce comprehensive microarrays of carbohydrates, glycoproteins and proteoglycans using isolated samples or cell extracts. Using standard microarray equipment, a series of carbohydrate microarrays were generated and probed with a panel of monoclonal antibodies with specificities for glycan epitopes. The arrays were highly reproducible, stable, and could be stored dry for several months. Glycans play central roles in development, carcinogenesis, cell adhesion, and immunity and are increasingly the subject of therapeutic approaches. We anticipate that the development of carbohydrate microarrays will be important for the high-throughput analysis of glycans and their molecular interactions.
DNA和蛋白质微阵列的发展代表了转录组学和蛋白质组学研究的重大进展。此类阵列允许对蛋白质的存在、相互作用以及基因表达进行高通量的平行分析。然而，在糖组分析方面，尚未有与之相当的技术与之匹配。原因之一是，与蛋白质相比，要可靠地固定化学和结构多样的聚糖群体较为困难。我们描述了一种新型微阵列载玻片表面的开发，在该表面上，无需对载玻片表面进行预先衍生化处理或对阵列样本进行任何修饰，就可以直接固定各种聚糖结构。这些载玻片可用于使用分离样本或细胞提取物制作碳水化合物、糖蛋白和蛋白聚糖的综合微阵列。利用标准的微阵列设备，生成了一系列碳水化合物微阵列，并用一组对聚糖表位具有特异性的单克隆抗体进行检测。这些阵列具有高度的可重复性、稳定性，并且可以干燥保存数月。聚糖在发育、致癌作用、细胞黏附及免疫过程中发挥着核心作用，并且越来越成为治疗方法的研究对象。我们预计碳水化合物微阵列的开发对于聚糖及其分子相互作用的高通量分析将具有重要意义。
