affinitykromatografia

Affinity chromatography is a separation technique used in biochemistry and molecular biology to purify a target molecule from a complex mixture. It relies on the specific binding interaction between the target molecule and a ligand immobilized on a stationary phase. This ligand is chosen for its ability to selectively bind to the molecule of interest, such as an antibody binding to an antigen, an enzyme binding to its substrate, or a receptor binding to its ligand.

The process begins by passing the mixture containing the target molecule through a chromatography column packed with the stationary phase. When the target molecule encounters the immobilized ligand, it binds specifically, while other components of the mixture that do not have affinity for the ligand pass through the column. After the unbound material has been washed away, the bound target molecule is eluted from the column by disrupting the specific binding interaction. This disruption can be achieved by changing the pH, ionic strength, or by adding a competing molecule that displaces the target molecule from the ligand.

Affinity chromatography is highly selective and can achieve significant purification in a single step. It is widely used for purifying proteins, antibodies, nucleic acids, and other biomolecules. The choice of ligand and stationary phase is crucial for the success of the separation and depends entirely on the specific target molecule being purified. Applications range from laboratory-scale research to large-scale industrial purification of biopharmaceuticals.