Flow field-flow fractionation: recent trends in protein analysis

Area of Science:

• Biochemistry
• Analytical Chemistry
• Separation Science

Background:

• Flow field-flow fractionation (F4) is a flow-assisted separation technique.
• It utilizes an empty channel and a perpendicular mobile phase flow.
• F4 separates analytes under native conditions without structural modification.

Purpose of the Study:

• To review recent literature on F4 applications for protein analysis.
• To highlight F4's advantages in separating biological macromolecules.
• To discuss F4's role in complementing established separation techniques.

Main Methods:

• Utilizing an empty channel as the separation device.
• Employing a second mobile phase flow as a perpendicular field.
• Analyzing intact proteins, protein complexes, oligomers, and aggregates.

Main Results:

• F4 enables gentle separation of biological samples, preserving original structure.
• The technique is increasingly applied to very high-molecular-weight protein species.
• F4 complements techniques like liquid chromatography and electrophoresis.

Conclusions:

• F4 is a promising technique for quality control of protein therapeutics.
• It is valuable for characterizing amyloid proteins and lipoprotein profiling.
• F4 serves as an effective pre-mass spectrometry separation step in proteomics.