Carbohydrate monolayer strategy for electrochemical assay of cell surface carbohydrate
Carbohydrate monolayer strategy for electrochemical assay of cell surface carbohydrate
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用于细胞表面碳水化合物电化学测定的碳水化合物单层策略
DOI：
10.1021/ja801468b
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发表时间：
2008-06-11
影响因子：
15
通讯作者：
Ju, Huangxian
中科院分区：
文献类型：
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作者：
Ding, Lin;Cheng, Wei;Ju, Huangxian
由于缺乏理想的检测工具，糖生物学的研究受到严重阻碍。这项工作提出了一个强大的碳水化合物单层平台，以解决与蛋白质阵列检测细胞表面碳水化合物报告的活性位点不可及性和凝集素变性的问题，并开发了一种方便的方法，用于监测细胞表面碳水化合物的网站的利益，具有高灵敏度，可接受的速度，低成本，和良好的可扩展性。它利用固体表面限制和细胞表面驻留的碳水化合物与量子点标记的碳水化合物识别蛋白的竞争性结合，以及随后的金属特征的伏安定量。甘露聚糖单层策略对K562细胞具有敏感的反应，并且由于凝集素与相应的碳水化合物之间的特异性相互作用而具有潜在的特异性。通过比较K562细胞与溶液中甘露聚糖的竞争性结合，可以估计单个K562细胞的平均Con A结合能力对应于6.9 pg或2.3 x 10（10）甘露糖部分。该策略综合了表面组装、纳米技术、生物偶联技术和电化学检测等技术的优点，由于量子点的多重编码能力，可以同时使用更多对凝集素和碳水化合物，从而扩展用于细胞表面碳水化合物的分析和高通量多重检测，为细胞表面碳水化合物位点的定量评价提供了重要的方法。
The study of glycobiology has been seriously hampered due to lack of an ideal assay tool. This work proposes a robust carbohydrate monolayer platform to solve the problems of active site inaccessibility and lectin denaturation associated with protein arrays reported for detection of cell surface carbohydrates and develops a convenient method for monitoring cell surface carbohydrate sites of interest, with high sensitivity, acceptable rapidity, low cost, and excellent extensibility. It utilizes the competitive binding of solid-surface-confined and cell-surface-residing carbohydrates to quantum dot labeled carbohydrate recognition protein and subsequent voltammetric quantification of the metal signature. The mannan monolayer strategy exhibited sensitive response to K562 cells and possessed potential specificity due to the specific interaction between lectin and corresponding carbohydrate. By comparing the competitive binding of K562 cells with mannan in solutions, the average Con A binding capacity of a single K562 cell could be estimated to correspond to 6.9 pg or 2.3 x 10(10) mannose moieties. This strategy integrates the advantages of surface assembly, nanotechnology, bioconjugate techniques, and electrochemical detection and can be expanded for profiling cell surface carbohydrates and high-throughput multiple detection by simultaneously using more pairs of lectin and carbohydrate owing to the multiple coding capability of QDs, which provides an important protocol for the quantitative evaluation of cell surface carbohydrate sites.