A versatile method for functionalizing surfaces with bioactive glycans.
A versatile method for functionalizing surfaces with bioactive glycans.
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DOI：
10.1021/bc1003372
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发表时间：
2011-01-19
影响因子：
4.7
通讯作者：
Ratner, Daniel M.
中科院分区：
文献类型：
--
作者：
Cheng, Fang;Shang, Jing;Ratner, Daniel M.
微阵列和生物传感器的功能归功于我们能够显示保留生物功能的表面结合的生物分子。用于将生物活性化合物固定在表面上的通用、稳定和简便的方法已经扩展了非专家实验室对分子相互作用的高通量“组学”规模筛选的应用。在此，我们证明了简化的化学物质的潜力，制造聚糖微阵列，利用二乙烯砜（DVS）修饰的表面共价固定的天然和化学衍生的碳水化合物，以及糖蛋白。通过表面等离子体共振成像（SPRi）定量检查捕获的聚糖的生物活性。通过X射线光电子能谱（XPS）和飞行时间二次离子质谱（ToF-SIMS）分别获得了DVS改性表面上碳水化合物物种的组成和光谱证据。基于相对亲核性（还原糖与胺和巯基衍生的糖苷）和端基异构体构型的聚糖的位点选择性固定也进行了研究。我们的研究结果表明，直接和可重复的共轭的各种功能的生物分子到乙烯基砜改性的生物传感器表面。这种方法的简单性将对糖组学研究产生重大影响，因为它扩展了非合成实验室快速构建功能性聚糖微阵列和定量生物传感器的能力。
Microarrays and biosensors owe their functionality to our ability to display surface-bound biomolecules with retained biological function. Versatile, stable, and facile methods for the immobilization of bioactive compounds on surfaces have expanded the application of high-throughput ‘omics’-scale screening of molecular interactions by non-expert laboratories. Herein, we demonstrate the potential of simplified chemistries to fabricate a glycan microarray, utilizing divinyl sulfone (DVS)-modified surfaces for the covalent immobilization of natural and chemically derived carbohydrates, as well as glycoproteins. The bioactivity of the captured glycans was quantitatively examined by surface plasmon resonance imaging (SPRi). Composition and spectroscopic evidence of carbohydrate species on the DVS-modified surface were obtained by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS), respectively. The site-selective immobilization of glycans based on relative nucleophilicity (reducing sugar vs. amine- and sulfhydryl-derived saccharides) and anomeric configuration was also examined. Our results demonstrate straightforward and reproducible conjugation of a variety of functional biomolecules onto a vinyl sulfone-modified biosensor surface. The simplicity of this method will have a significant impact on glycomics research, as it expands the ability of non-synthetic laboratories to rapidly construct functional glycan microarrays and quantitative biosensors.
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