FPLC

Fast Protein Liquid Chromatography, often abbreviated as FPLC, is a technique used for the separation and purification of biomolecules such as proteins, peptides, and nucleic acids. It is a type of liquid chromatography that operates at moderate pressures, distinguishing it from High-Performance Liquid Chromatography (HPLC). FPLC systems are designed for high resolution and recovery of biological samples, making them ideal for preparative and analytical purposes in biochemistry, molecular biology, and pharmaceutical research.

The FPLC process involves a mobile phase (a buffer solution) that is pumped through a stationary phase packed within a chromatography column. The stationary phase is typically a resin with specific properties that interact with the target molecules. These interactions can be based on various principles, including size exclusion, ion exchange, affinity, or hydrophobic interactions. As the sample is introduced and carried through the column by the mobile phase, different components interact with the stationary phase to varying degrees. Molecules that interact less strongly with the stationary phase will elute (exit the column) earlier, while those that interact more strongly will be retained longer.

FPLC systems typically consist of a pump, an injector, a column, and a detector. The pump delivers the mobile phase at a controlled flow rate. The injector introduces the sample into the mobile phase stream. The column houses the stationary phase where separation occurs. The detector monitors the eluent and generates a chromatogram, a graphical representation of the separation, which shows peaks corresponding to different eluted components. FPLC is widely used for protein purification, analysis of protein-protein interactions, and quality control of biopharmaceuticals.