Affinity chromatography in which a macromolecule, (for example, a protein) is separated and purified in a solution with a molecule containing a ligand for which the macromolecule has an affinity that reason it is retained on the column.
The Advantages of Affinity Chromatography are:
- High sensitivity compared to TCD & FID.
- Affinity chromatography is used in the production of vaccines.
- Affinity chromatography is used in the purification of protein and enzyme.
- Affinity chromatography gives High specificity.
- Enzymes and other proteins are studied by affinity chromatography.
- To maintain the quality of the product, this chromatography is used in the pharmaceutical manufacturer in the production of vaccines.
- Affinity chromatography doesn’t rely on ionic strength, pH, temperature, and composition of the buffer.
- The high degree of purity can be obtained by Affinity Chromatography.
- This is a very reproducible process.
- This is the simplification method.
- Used to increase the solubility.
- In genetic engineering, affinity chromatography is used.
- It takes a lot of skill to handle it.
- It interferes with the structure.
- Transfer and the leakage of metal ion lead to protein loss.
- Sometimes ligands leakage is observed.
- The volume of the sample is limited.
- The carrier gas used must be pure such as pure nitrogen.
- The ligands used in affinity chromatography are costly.
- Relatively low productivity.
- It has a non-specific adsorption.
- Degradation of the solid support.
- Metal-ion transfer and metal ion leakage lead to loss of protein.
- Affinity chromatography is non-specific for adsorption than other chromatography methods.
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