Haab Brian B, Partyka Katie, Cao Zheng
Van Andel Research Institute, Grand Rapids, Michigan.
Curr Protoc Protein Sci. 2013 Sep 24;73:27.6.1-27.6.16. doi: 10.1002/0471140864.ps2706s73.
Antibody arrays provide a valuable method for obtaining multiple protein measurements from small volumes of biological samples. Antibody arrays can be designed to target not only core protein abundances (relative or absolute abundances, depending on the availability of standards for calibration), but also posttranslational modifications, provided antibodies or other affinity reagents are available to detect them. Glycosylation is a common modification that has important and diverse functions in both normal and disease biology. Significant progress has been made in developing methods for measuring glycan levels on multiple specific proteins using antibody arrays and glycan-binding reagents. This unit describes practical approaches for developing, optimizing, and using antibody array assays to determine both protein abundance and glycosylation state. Low-volume arrays can be used to reduce sample consumption, and a new way to improve the binding strength of particular glycan-binding reagents through multimerization is discussed. These methods can be useful for a wide range of biological studies in which glycosylation may change and/or affect protein function.
抗体阵列提供了一种从少量生物样品中获取多种蛋白质测量值的宝贵方法。抗体阵列不仅可以设计用于靶向核心蛋白质丰度（相对或绝对丰度，取决于校准标准的可用性），还可以用于检测翻译后修饰，前提是有可用的抗体或其他亲和试剂来检测它们。糖基化是一种常见的修饰，在正常生物学和疾病生物学中都具有重要且多样的功能。在开发使用抗体阵列和聚糖结合试剂测量多种特定蛋白质上聚糖水平的方法方面已经取得了重大进展。本单元描述了开发、优化和使用抗体阵列分析来确定蛋白质丰度和糖基化状态的实用方法。低体积阵列可用于减少样品消耗，并讨论了一种通过多聚化提高特定聚糖结合试剂结合强度的新方法。这些方法可用于广泛的生物学研究，其中糖基化可能会发生变化和/或影响蛋白质功能。
