what is hplc

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.

The Disadvantages of Affinity Chromatography are:

  • 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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