快速蛋白质液相色谱法

Fast Protein Liquid Chromatography.

作者信息

Madadlou Ashkan, O'Sullivan Siobhan, Sheehan David

机构信息

Tehran University, Food Science and Engineering, Karadj, Iran.

Department of Biological Sciences, Cork Institute of Technology, Cork, Ireland.

出版信息

Methods Mol Biol. 2017;1485:365-373. doi: 10.1007/978-1-4939-6412-3_19.

DOI:10.1007/978-1-4939-6412-3_19

PMID:27730563

Abstract

Fast protein liquid chromatography (FPLC) is a form of high-performance chromatography that takes advantage of the high resolution made possible by small-diameter stationary phases. It was originally developed for proteins and features high loading capacity, biocompatible aqueous buffer systems, fast flow rates, and availability of stationary phases in most common chromatography modes (e.g., ion exchange, gel filtration, reversed phase, and affinity). The system makes reproducible separation possible by incorporating a high level of automation including autosamplers, gradient program control, and peak collection. In addition to proteins, the method is applicable to other kinds of biological samples including oligonucleotides and plasmids. The most common type of FPLC experiment is anion exchange of proteins. This chapter describes such an experiment carried out using an ÄKTA FPLC explorer system (Amersham Pharmacia Biotech, Sweden).

摘要

快速蛋白质液相色谱（FPLC）是一种高效色谱形式，它利用小直径固定相实现高分辨率。它最初是为蛋白质开发的，具有高负载能力、生物相容性水性缓冲系统、快速流速以及大多数常见色谱模式（如离子交换、凝胶过滤、反相和亲和）的固定相可用性。该系统通过纳入包括自动进样器、梯度程序控制和峰收集在内的高水平自动化，实现了可重复的分离。除蛋白质外，该方法还适用于其他种类的生物样品，包括寡核苷酸和质粒。最常见的FPLC实验类型是蛋白质的阴离子交换。本章描述了使用ÄKTA FPLC探索者系统（瑞典阿默沙姆 Pharmacia生物技术公司）进行的此类实验。