N-Glycan Analysis by Ultra-Performance Liquid Chromatography and Capillary Gel Electrophoresis with Fluorescent Labeling

Q1 Biochemistry, Genetics and Molecular Biology

Current Protocols in Protein Science Pub Date : 2019-09-04 DOI:10.1002/cpps.95

Maja Hanić, Gordan Lauc, Irena Trbojević-Akmačić

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引用次数: 12

Abstract

Glycans are a class of macromolecules essential for all forms of life. They embellish various proteins and other macromolecules in organisms and are responsible for their proper functioning. Because their complex structure is determined by genetic and environmental factors, analysis of such molecules is rather demanding. Liquid chromatography (high-performance and ultra-performance, HPLC and UPLC, respectively) analysis has been used for the purpose of glycoprofiling for years and it is a well-established method regarding its robustness, reproducibility, and high throughput. Another orthogonal method that is now used in glycoprofiling is capillary gel electrophoresis (CGE) because it offers powerful separation and distinct sensitivity. The purpose of the following protocols is to present all steps required for release and fluorescent labeling of total N-glycans from blood plasma/serum or isolated glycoprotein (for example, IgG) and their subsequent UPLC or CGE analysis. © 2019 by John Wiley & Sons, Inc.

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n -聚糖的超高效液相色谱和荧光标记毛细管凝胶电泳分析

聚糖是所有生命形式所必需的一类大分子。它们修饰生物体中的各种蛋白质和其他大分子，并负责它们的正常功能。由于其复杂的结构是由遗传和环境因素决定的，对这类分子的分析要求相当高。液相色谱(分别为高效液相色谱和超高效液相色谱)分析已被用于糖谱分析多年，它是一种成熟的方法，具有稳健性、重复性和高通量。另一种用于糖谱分析的正交方法是毛细管凝胶电泳(CGE)，因为它具有强大的分离能力和独特的灵敏度。以下方案的目的是介绍从血浆/血清或分离的糖蛋白(例如IgG)中释放和荧光标记总n -聚糖及其随后的UPLC或CGE分析所需的所有步骤。©2019 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Current Protocols in Protein Science Biochemistry, Genetics and Molecular Biology-Biochemistry

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期刊介绍： With the mapping of the human genome, more and more researchers are exploring protein structures and functions in living organisms. Current Protocols in Protein Science provides protein scientists, biochemists, molecular biologists, geneticists, and others with the first comprehensive suite of protocols for this growing field.