Pilobello Kanoelani T, Agrawal Praveen, Rouse Richard, Mahal Lara K
Department of Chemistry, Biomedical Chemistry Institute, New York University, New York, USA.
Curr Protoc Chem Biol. 2013;5(1):1-23. doi: 10.1002/9780470559277.ch120035.
Lectin microarray technology has been used to profile the glycosylation of a multitude of biological and clinical samples, leading to new clinical biomarkers and advances in glycobiology. Lectin microarrays, which include >90 plant lectins, recombinant lectins, and selected antibodies, are used to profile N-linked, O-linked, and glycolipid glycans. The specificity and depth of glycan profiling depends upon the carbohydrate-binding proteins arrayed. The current set targets mammalian carbohydrates including fucose, high mannose, branched and complex N-linked, α- and β-galactose and GalNAc, α-2,3- and α-2,6-sialic acid, LacNAc, and Lewis X epitopes. Previous protocols have described the use of a contact microarray printer for lectin microarray production. Here, an updated protocol that uses a non-contact, piezoelectric printer, which leads to increased lectin activity on the array, is presented. Optimization of print and sample hybridization conditions and methods of analysis are discussed.
凝集素微阵列技术已被用于分析大量生物和临床样本的糖基化情况，从而产生了新的临床生物标志物，并推动了糖生物学的发展。凝集素微阵列包含90多种植物凝集素、重组凝集素和选定的抗体，用于分析N-连接、O-连接和糖脂聚糖。聚糖分析的特异性和深度取决于所排列的碳水化合物结合蛋白。当前的阵列针对哺乳动物碳水化合物，包括岩藻糖、高甘露糖、分支和复杂的N-连接糖、α-和β-半乳糖以及N-乙酰半乳糖胺、α-2,3-和α-2,6-唾液酸、乳糖胺和Lewis X表位。以前的方案描述了使用接触式微阵列打印机生产凝集素微阵列。在此，我们提出了一种更新的方案，该方案使用非接触式压电打印机，可提高阵列上凝集素的活性。文中还讨论了打印和样本杂交条件的优化以及分析方法。
