Fast protein liquid chromatography

Fast Protein Liquid Chromatography, usually referred to as FPLC, is a form of column chromatography used to separate or purify proteins from complex mixtures by using Ion Chromatography. It is very commonly used in biochemistry and enzymology. Columns used with an FPLC can separate macromolecules based on size, charge distribution, hydrophobicity, or biorecognition (as with affinity chromatography).

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Trademark

FPLC is a trademark of GE Healthcare, formerly Amersham-Pharmacia Biotech and Pharmacia Biotech.

Typical columns

Typical columns used for protein purification include:

• Ion exchange chromatography, cation or anion — separate proteins based on surface-charges
• Size exclusion chromatography, also known as Gel filtration chromatography, Gel permeation chromatography or desalting — separate proteins based on size.
• Reversed phase or hydrophobic interaction — separates based on hydrophobicity which depends heavily on surface area
• Affinity chromatography — columns which purify based on ligand affinity (e.g. a His₆-tagged protein purified on a nickel column).

See also

• Chromatography, an overview article covering all chromatographic techniques.
• High performance liquid chromatography (HPLC) for column chromatography using high pressure.
• Affinity chromatography