Novel 3-dimensional dendrimer platform for glycolipid microarray.
Novel 3-dimensional dendrimer platform for glycolipid microarray.
复制标题
用于糖脂微阵列的新型 3 维树枝状聚合物平台。
DOI：
10.1016/j.bios.2011.07.044
复制
发表时间：
2011
影响因子：
12.6
通讯作者：
Zhou,Xichun
中科院分区：
文献类型：
--
作者：
Zhang,Jian;Zhou,Xichun
糖脂是调节细胞识别和病原体黏附的重要生物分子。在这篇文章中，我们报道了一种灵敏的糖脂微阵列，用于在微阵列底物上非共价固定糖脂，并进行了一系列免疫分析来探索糖脂与蛋白质的相互作用。这种衬底利用了附着在微观玻璃表面的三维酰肼功能化树枝状大分子单分子膜，它具有以非共价方式吸附糖链的特性，并促进了衬底表面的多价属性。在概念验证实验中，在底物上检测神经节苷脂GM1、FucGM1、GM3、GD1b、GT1b和GQ1b，以及一种脂阿拉伯甘露聚糖，并用毒素和抗体进行询问。由此得到的糖脂微阵列在检测糖脂-蛋白质相互作用方面表现出超敏和特异性。特别是，五聚体霍乱毒素B亚单位与阵列上斑点的GM1糖脂的牢固和特异结合表明了其在敏感性和特异性方面的优越性。此外，该糖脂微阵列底物用于检测缓冲液中的阿拉伯甘露聚糖，检出限为125 ng/mL。此外，在该平台上对人尿标本进行了结核分枝杆菌(Mtb)阿拉伯甘露聚糖的检测，该方法可以有效地识别感染或未感染结核分枝杆菌的尿样。这项工作的结果表明，利用该糖脂微阵列平台制备包括游离糖和糖脂在内的糖共轭微阵列的可能性，以及在病原体和毒素检测中的潜在应用。
Glycolipids are important biological molecules that modulate cellular recognitions and pathogen adhesions. In this paper, we report a sensitive glycolipid microarray for non-covalently immobilizing glycolipids on a microarray substrate and we perform a set of immunoassays to explore glycolipid–protein interactions. This substrate utilizes a three-dimensional hydrazide-functionalized dendrimer monolayer attached onto a microscopic glass surface, which possesses the characteristics to adsorb glycoliplids non-covalently and facilitates multivalent attributes on the substrate surface. In the proof-of-concept experiments, gangliosides such as GM1, FucGM1, GM3, GD1b, GT1b, and GQ1b, and a lipoarabinomannan were tested on the substrate and interrogated with toxins and antibodies. The resulting glycolipid microarrays exhibited hypersensitivity and specificity for detection of glycolipid–protein interactions. In particular, a robust and specific binding of a pentameric cholera toxin B subunit to the GM1 glycolipid spotted on the array has demonstrated its superiority in sensitivity and specificity. In addition, this glycolipid microarray substrate was used to detect lipoarabinomannan in buffer within a limit-of-detection of 125ng/mL. Furthermore, Mycobacterium tuberculosis (Mtb) Lipoarabinomannan was tested in human urine specimens on this platform, which can effectively identify urine samples either infected or not infected with Mtb. The results of this work suggest the possibility of using this glycolipid microarray platform to fabricate glycoconjugate microarrays, which includes free glycans and glycolipids and potential application in detection of pathogen and toxin.