The following are several commonly used monoclonal antibody purification methods based on progressive purification:
1. Affinity chromatography: Use antibody-specific recognition to separate antibodies. Adsorbents such as protein A, protein G or protein L are usually used to perform affinity chromatography on the common heavy and light chains, Fc regions and other parts of antibodies.
2. Ion exchange chromatography: Use the surface-charged residues in the molecule to interact with the oppositely charged ion exchange matrix to specifically purify the target molecule. The selection can be based on the isoelectric point of the antibody.
3. Gel filtration chromatography: Use pore size to selectively separate antibodies and other molecules to remove macromolecular impurities.
4. Reversed-phase chromatography: According to the different hydrophilicity of target molecules and contaminant molecules in the solution, electrostatic force is used to separate antibodies. Reversed-phase chromatography has the advantage of being insensitive to ionic strength and is widely used in the purification process of antibodies.
The choice of monoclonal antibody purification method will be affected by many factors, such as mother solution concentration, cation contamination, pH value, etc.