Fast protein liquid chromatography (FPLC) is a form of medium-pressure chromatography that uses a pump to control the speed at which the mobile phase passes through the stationary phase. FPLC was introduced in 1982 by Pharmacia as fast performance liquid chromatography. Since then, many different medium-pressure chromatography systems have been developed. It should be noted that researchers often use the terms fast protein liquid chromatography, FPLC, and medium-pressure chromatography interchangeably.
Related Topics: Types of Chromatography, Liquid Chromatography Principles, Column Chromatography Methods and Instrumentation
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Fast protein liquid chromatography systems generally consist of a pump, a UV detector, a conductivity meter, and a fraction collector and operate at pressures of ~3,500 psi (24 MPa). Some fast protein liquid chromatography systems also have column switching valves that allow the user to switch between columns, sample pumps to apply the sample to a column, and buffer blending valves that can generate desired buffer gradients. Modern fast protein liquid chromatography systems such as the NGC™ medium-pressure chromatography system enable most user configuration and method creation via a dedicated software interface.
Fig. 1. Sample fast protein liquid chromatography flow path. A sample flow path that illustrates the components of the NGC medium-pressure chromatography system. Image is taken from the NGC system’s ChromLab™ software.
Samples can either be loaded manually by injection into a sample loop or automatically using a sample pump. Some fast protein liquid chromatography systems have multi-wavelength detectors for monitoring sample elution at several wavelengths. This feature can be particularly useful when working with chromogenic or fluorescently tagged proteins that absorb at wavelengths other than 280 nm, as elution of the protein of interest can be tracked separately from elution of contaminating proteins based on differences in absorption.
For a more detailed description of fast protein liquid chromatography systems, visit our Medium-Pressure Chromatography page.