It is also used to collect separated compounds of interest and identify protein profiles within single Biomolecule mixtures can include proteins, peptides, oligonucleotides, DNA, and RNA. Because biomolecules are highly sensitive and can’t withstand the type of high pressure, temperature, or solvents used in high-performance liquid chromatography (HPLC), FPLC is employed. FPLC is also used to collect separated compounds of interest and identify protein profiles within single proteins.

You may hear FPLC referred to as medium-pressure chromatography, biochromatography, or biopurification, but the title doesn’t change the process. It’s an important method for many researchers in the life sciences and biotech.

Separating a sample within a mixture depends on each molecule’s specific interaction with the stationary and mobile phases used during chromatography. FPLC utilizes a wide range of resins, composed of many small coated beads, for its stationary phase, and a an aqueous solution or buffer for its mobile phase.

The targeted protein adheres to the resin as it flows through the separation column and the buffer carries any other molecules that are present out of the system. It’s typical to see a mixture of buffers used to accomplish this elution.

For example, having flushed the system, a secondary buffer, called an eluent, will separate the protein of interest from the resin and pull it into a detection system for measurement, recording the concentration levels of the target protein in the sample.

FPLC is a type of preparative technique, unlike HPLC which provides quantitative and qualitative analysis of liquid samples. It is used in all sorts of settings, from academia to research facilities to industrial manufacturing.

Its versatility is due in part to its simplicity, rapidity, and reliability as a technique, producing consistent results that end-users can count on.

Furthermore, its ability to support multiple columns simultaneously is based on its use of low pressure and offers a wide flow range when combined with multiple pumps, making the entire process efficient and highly scalable.

Some of the most common FPLC systems on the market include the AKTA FPLC system, offered by Cytiva—formerly GE Healthcare and Amersham Biosciences—and the AZURA® FPLC system, offered by KNAUER. (And guess what? We lease them.)