Multiplexed analysis of glycan variation on native proteins captured by antibody microarrays
Multiplexed analysis of glycan variation on native proteins captured by antibody microarrays
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DOI：
10.1038/nmeth1035
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发表时间：
2007-05-01
期刊：
影响因子：
48
通讯作者：
Haab, Brian B.
中科院分区：
文献类型：
--
作者：
Chen, Songming;LaRoche, Tom;Haab, Brian B.
蛋白质上的碳水化合物翻译后修饰是正常和疾病生物学中蛋白质功能的重要决定因素。我们开发了一种方法，可以使用抗体微阵列捕获多种蛋白质，然后用凝集素或聚糖结合抗体进行检测，从而对复杂混合物中单个蛋白质的聚糖进行高效、多重研究。斑点抗体上聚糖的化学衍生化阻止了凝集素与这些聚糖的结合。多种凝集素可用作检测探针，每种探针针对不同的聚糖基团，以建立捕获蛋白质的凝集素结合谱。通过使用平行夹心和聚糖检测分析分析多个样品中的蛋白质和聚糖变异，我们发现胰腺癌患者血清中蛋白质 MUC1 和 CEA 上存在与癌症相关的聚糖变化。用于聚糖检测的抗体阵列对于分析多种蛋白质上特定聚糖的变异非常有效，并且应该在糖生物学研究的各个领域都有用。
Carbohydrate post-translational modifications on proteins are important determinants of protein function in both normal and disease biology. We have developed a method to allow the efficient, multiplexed study of glycans on individual proteins from complex mixtures, using antibody microarray capture of multiple proteins followed by detection with lectins or glycan-binding antibodies. Chemical derivatization of the glycans on the spotted antibodies prevented lectin binding to those glycans. Multiple lectins could be used as detection probes, each targeting different glycan groups, to build up lectin binding profiles of captured proteins. By profiling both protein and glycan variation in multiple samples using parallel sandwich and glycan-detection assays, we found cancer-associated glycan alteration on the proteins MUC1 and CEA in the serum of pancreatic cancer patients. Antibody arrays for glycan detection are highly effective for profiling variation in specific glycans on multiple proteins and should be useful in diverse areas of glycobiology research.