Surface Plasmon Resonance Imaging Analysis of Protein Binding to a Sialoside-Based Carbohydrate Microarray
Surface Plasmon Resonance Imaging Analysis of Protein Binding to a Sialoside-Based Carbohydrate Microarray
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DOI：
10.1007/978-1-61779-373-8_13
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Cheng, Quan
中科院分区：
文献类型：
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作者：
Linman, Matthew J.;Yu, Hai;Cheng, Quan
以多路复用阵列格式监测多种生物相互作用具有许多优点。然而，将成熟的表面化学用于光谱测量转化为基于阵列的高通量筛选并不是一个简单的过程，并且经常被证明是方法开发的瓶颈。本章报道了一种新的碳水化合物微阵列的制备和表征，该微阵列具有合成的硅酸苷，用于表面等离子体共振成像分析凝集素碳水化合物的相互作用。采用接触印刷技术在中性皂素包被表面上印制功能性皂苷，并用荧光显微镜对所得元素的性质进行了表征。用黑参凝集素（SNA）在四种不同的糖功能化表面进行了检测，并分析了其结合差异。SNA/生物素化涎苷相互作用的多重检测在多达400个元素的阵列上进行了良好的数据相关性，证明了生物素-中性蛋白为基础的生物界面在控制探针方向以实现可重复和有效的蛋白质与碳水化合物结合方面的有效性。
Monitoring multiple biological interactions in a multiplexed array format has numerous advantages. However, converting well-developed surface chemistry for spectroscopic measurements to array-based, high-throughput screening is not a trivial process and often proves to be the bottleneck in method development. This chapter reports the fabrication and characterization of a new carbohydrate microarray with synthetic sialosides for surface plasmon resonance imaging analysis of lectin carbohydrate interactions. Contact printing of functional sialosides on neutravidin-coated surfaces was carried out and the properties of the resulting elements were characterized by fluorescence microscopy. Sambucus nigra agglutinin (SNA) was used for testing on four different carbohydrate-functionalized surfaces and differential binding was analyzed. Multiplexed detection of SNA/biotinylated sialoside interactions on arrays up to 400 elements has been performed with good data correlation, demonstrating the effectiveness or the biotin-neutravidin-based biointerface to control probe orientation for reproducible and efficient protein binding to carbohydrates.