Applications, Advantages & Disadvantages of Affinity chromatography

Applications:

1. Protein Purification:

• Widely used to purify proteins from complex mixtures, especially recombinant proteins tagged for easier isolation.

2. Enzyme Isolation:

• Retrieve specific enzymes from cellular extracts.

3. Drug Target Identification:

• Used in drug discovery to identify potential cellular targets of a new drug.

4. Antibody Purification:

• Immunoaffinity chromatography for isolating specific antibodies.

5. Removal of Specific Contaminants:

• For example, endotoxin removal from biopharmaceutical preparations.

Here's a diagram illustrating the applications of Affinity Chromatography:

Advantages:

1. High Specificity:

• Thanks to the specific ligand-target molecule interaction.

2. High Purity:

• Achieve high purity levels in a single chromatographic step.

3. Versatility:

• A wide range of ligands can be immobilized, making it suitable for various applications.

4. Scalability:

From micrograms in research labs to kilograms in the industry.

Here's a diagram illustrating the advantages of Affinity Chromatography:

Disadvantages:

1. Ligand Leakage:

• The ligand can sometimes leach from the column, leading to contamination.

2. Slow Process:

• Due to the necessity for equilibrium during binding, the process can be time-consuming.

3. Expense:

• High costs associated with specialized affinity ligands.

4. Non-specific Binding:

• Despite the specificity, some non-target molecules might also bind to the column, leading to impurities.

Here's a diagram illustrating the disadvantages of Affinity Chromatography: