A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates
A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates
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DOI：
10.1021/ja0613963
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发表时间：
2006-08-16
影响因子：
15
通讯作者：
Hamachi, Itaru
中科院分区：
文献类型：
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作者：
Koshi, Yoichiro;Nakata, Eiji;Hamachi, Itaru
由于糖及其衍生物在各种生物现象中起着重要作用，因此迫切需要对各种糖缀合物进行快速和高通量的分析。我们在此描述了一种新的荧光凝集素阵列的构建，用于糖检测使用超分子水凝胶基质。在该阵列中，荧光凝集素在半湿条件下非共价固定以抑制蛋白质变性。荧光滴定和荧光寿命实验表明，固定化凝集素作为分子识别支架在水凝胶基质中，类似于在水溶液中。也就是说，双分子荧光猝灭和恢复（BFQR）方法可以在两种条件下成功地操作。这使得人们能够基于固定化凝集素的识别选择性和亲和性荧光读出一系列荧光，而不需要繁琐的洗涤过程，也不需要标记目标荧光。使用本凝集素阵列对不同的糖缀合物进行简单和高通量的传感和分析，所述糖缀合物不仅包括简单的葡萄糖，还包括寡糖和糖蛋白，并且，此外，还完成了几种类型的细胞裂解物的模式识别和分析。
Because sugar and its derivatives play important roles in various biological phenomena, the rapid and high-throughput analysis of various glycoconjugates is keenly desirable. We describe herein the construction of a novel fluorescent lectin array for saccharide detection using a supramolecular hydrogel matrix. In this array, the fluorescent lectins were noncovalently fixed under semi-wet conditions to suppress the protein denaturation. It is demonstrated by fluorescence titration and fluorescence lifetime experiments that the immobilized lectins act as a molecular recognition scaffold in the hydrogel matrix, similar to that in aqueous solution. That is, a bimolecular fluorescence quenching and recovery (BFQR) method can successfully operate under both conditions. This enables one to fluorescently read-out a series of saccharides on the basis of the recognition selectivity and affinity of the immobilized lectins without tedious washing processes and without labeling the target saccharides. Simple and high-throughput sensing and profiling were carried out using the present lectin array for diverse glycoconjugates, which not only included a simple glucose, but also oligosaccharides, and glycoproteins, and, furthermore, the pattern recognition and profiling of several types of cell lysates were also accomplished.