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Affinity Chromatography

By

Dr. Mehboob Peeran

BIOLOGICALLY ACTIVE MOLECULES THAT HAVE BEEN PURIFIED BY THIS TECHNIQUE Enzymes, hormones, drug receptors, Vitamins, or cofactor binding proteins, antibodies, sulphahydryl group containing proteins.

Specific examples
Ligand
Protein(using Sepharose)

ADVANTAGES

1. Single step purification

2. Very few manipulations

3. The matrix can be used repeatedly

4. The matrix is a solid can be easily washed and dried

5. Research potential and industrial applications.

 

DISADVANTAGES

1. Since affinity is highly specific, conditions employed are also specific

2. Non specific adsorptions and elution possibilities exist. So standardisations of conditions is extremely important.

3. Polymer support is expensive.

4. Repeated use is always not realised.

5. Deactivation of the ligand, loss of activity of the purified sample is possible.

Acknowledgement

The author had attended a ten day workshop on the subject conducted by the Primate Research Lab in the Indian Institute of Science, Bangalore, several years ago. Most of the material in this article is from the lectures and information obtained from that workshop.

Copyrights: 2005 www.chemvista.org All Rights Reserved
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