Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples

Chen Lu; Joshua L. Wonsidler; Jianwei Li; Yanming Du; Timothy Block; Brian Haab; Songming Chen

doi:10.3791/3791

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Biology

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JoVE Journal Biology

Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples

化学阻断抗体芯片复用高通量的复杂样品中特定蛋白糖基化的剖析

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13:21 min

May 4, 2012

DOI: 10.3791/3791-v

Chen Lu¹, Joshua L. Wonsidler¹, Jianwei Li², Yanming Du¹, Timothy Block³, Brian Haab⁴, Songming Chen¹

¹Institute for Hepatitis and Virus Research, ²Department of Microbiology and Immunology,Thomas Jefferson University, ³Drexel University College of Medicine, ⁴Van Andel Research Institute

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在这项研究中，我们描述了一个复用高通量抗体芯片与外源凝集素的检测方法，可以在特定的蛋白质糖基化分析改进协议。该协议提供了新的可靠的试剂和显着降低，相比以前的程序的时间，成本和实验室设备的要求。

本视频文章介绍了使用化学封闭抗体微阵列获取肝细胞癌患者血清样品中 20 种不同糖蛋白的糖基化谱的程序：首先将糖蛋白特异性抗体直接打印到微阵列载玻片上，并封闭抗体聚糖以防止凝集素结合。当应用患者血清样品时，样品中的糖蛋白被抗体捕获。然后加入生物素化的聚糖结合蛋白以检测聚糖，并加入二偶联的 neu travain 以标记生物素化的凝集素。

然后扫描微阵列玻片以检测荧光，结果数据的分析揭示了样品蛋白质的糖基化谱。与 HPLC 等现有方法相比，该技术具有几个优点。该方法允许检测处于天然状态的单个糖蛋白。

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