Fast Protein Liquid Chromatography

Instrument: A wide range of low pressure LC systems are available, predominately AKTA-based systems from GE

Low pressure liquid chromatography is a rapid and robust method for purification of molecules from a mixture. The sample is applied to a column composed of functionalised beads to which the molecules in the sample can bind. Liquid is pumped through the column and, depending on the properties of the liquid relative to the beads, the sample molecules and any contaminants are sequentially removed.

Fast protein liquid chromatography is just liquid chromatography in which the beads are functionalised specifically to bind particular types of protein, such as those that carry a tag that binds metal ions. One of the most common types of protein chromatography is ion exchange, where positively charged proteins will bind to to negatively charged beads, or vice versa, and proteins are removed from the column by altering pH or ionic strength of the liquid. The output from the column is typically monitored by UV absorbance and collected in fractions.

Liquid chromatography is extremely widely used for preparative and semi-preparative separations and purifications of molecules ranging from small chemicals to large macromolecules.
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