Fast Protein Liquid Chromatography

Abstract

Fast protein liquid chromatography (FPLC) is a form of high-performance chromatography that takes advantage of 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).