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使用抗体微阵列分析血清蛋白上的聚糖。

Analysis of glycans on serum proteins using antibody microarrays.

作者信息

Chen Songming, Haab Brian B

机构信息

Van Andel Research Institute, Grand Rapids, MI, USA.

出版信息

Methods Mol Biol. 2009;520:39-58. doi: 10.1007/978-1-60327-811-9_4.

Abstract

Antibody arrays can be employed for the profiling glycan structures on proteins. Antibody arrays capture multiple, specific proteins directly from biological samples (such as serum), and lectin and glycan-binding antibodies probe the levels of specific glycans on the captured proteins. We use a practical method of partitioning microscope slides to enable the convenient processing of many detection reagents or samples. A critical first step in the procedure is the chemical derivatization of the glycans on the spotted capture antibodies, which prevents lectin binding to those glycans. We describe those methods along with the methods for preparing and treating serum samples, running the experiments, and designing and interpreting the experiments.

摘要

抗体阵列可用于分析蛋白质上的聚糖结构。抗体阵列可直接从生物样品(如血清)中捕获多种特定蛋白质,凝集素和聚糖结合抗体则可探测捕获蛋白质上特定聚糖的水平。我们采用一种实用的方法对显微镜载玻片进行分区,以便方便地处理许多检测试剂或样品。该过程的关键第一步是对斑点捕获抗体上的聚糖进行化学衍生化,这可防止凝集素与这些聚糖结合。我们将描述这些方法以及血清样品的制备和处理方法、进行实验的方法以及实验设计和解读方法。

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