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Lectin-Based Nanoprobes Functionalized with Enzyme for Highly Sensitive Electrochemical Monitoring of Dynamic Carbohydrate Expression on Living Cells

机译:酶功能化的基于凝集素的纳米探针,用于高灵敏度电化学监测活细胞中动态碳水化合物的表达

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摘要

A dual-functionalized nanoprobe was designed for highly sensitive and selective in situ evaluation of carbohydrates on living cells by integrating the specific carbohydrate recognition and enzymatic signal amplification of proteins on Au nanoparticles. A nanoscaffold of nanohorns functionalized with arginine-glycine-aspartic acid-serine tetrapeptide was also prepared on an electrode surface for cell capture and enhancing the electrical connectivity. Combined with the nanoprobe and peptide-nanohorns, a highly sensitive electrochemical strategy was developed for cytosensing, which showed a detection limit down to 15 cells, broad dynamic range, acceptable rapidity, and low cost. The proposed method was further used for monitoring of dynamic variation of carbohydrate expression on cancer cells in response to drugs, which obviated the destruction or labeling of cells and the covalent tagging of lectin and enzyme. The one-pot conjugation of three components was very convenient and could be extended for preparation of other lectin-functionalized nanoprobes. Further development of this technique would contribute considerably to meeting the challenges in comprehensive understanding of the glycomic codes.
机译:通过整合特异性的碳水化合物识别和Au纳米颗粒上蛋白质的酶促信号放大,设计了一种双功能纳米探针,用于对活细胞上的碳水化合物进行高度灵敏和选择性的原位评估。还在电极表面上制备了用精氨酸-甘氨酸-天冬氨酸-丝氨酸四肽官能化的纳米角纳米支架,用于细胞捕获和增强电连接性。结合纳米探针和肽纳米角,开发了一种用于细胞感测的高度敏感的电化学策略,该策略显示出检测限低至15个细胞,宽动态范围,可接受的快速度和低成本。所提出的方法还用于监测药物对癌细胞中碳水化合物表达的动态变化,从而避免了细胞的破坏或标记以及凝集素和酶的共价标记。一锅结合三种成分非常方便,可以扩展到其他凝集素功能化纳米探针的制备。该技术的进一步发展将大大有助于应对全面理解糖基密码的挑战。

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