Affinity Chromatography

Abstract

Affinity chromatography is a separation technique based on the use of specific and selective immobilized ligands able to associate reversibly to a desired biomolecule. Samples are passed through affinity columns that are equilibrated with a buffer favouring interaction between the immobilized ligand and the target biomolecule. Unrelated contaminants will be eluted in the column void volume and a suitable buffer change will lead to elution of the adsorbed biomolecule.

Keywords: molecular recognition; purification; immobilized ligands; derivatization

References

Chaiken IM (1987) Analytical Affinity Chromatography. Boca Raton. CRC Press.

Fassina G, Verdoliva A, Odierna MR, Ruvo M and Cassani G (1996) Protein A mimetic peptide ligand for affinity purification of antibodies. Journal of Molecular Recognition 9: 564–569.

Hjelm H, Hjelm K and Sjoquist J (1972) Protein A from Staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins. FEBS Letters 28: 73–76.

Nevens JR, Mallia AK, Wendt MW and Smith PK (1992) Affinity chromatographic purification of immunoglobulin M antibodies utilizing immobilized mannan binding protein. Journal of Chromatography 597: 247–256.

Roque‐Barreira MR and Campos‐Nieto A (1985) Jacalin: an IgA‐binding lectin. Journal of Biological Chemistry 134: 1740–1743.

Vretblad P (1976) Purification of lectins by biospecific affinity chromatography. Biochimica et Biophysica Acta 434: 169–176.

Wilson JE (1976) Application of Blue Dextran and Cibacron Blue F3GA in purification and structural studies of nucleotide requiring enzymes. Biochemical and Biophysical Research Communications 72: 816–823.

Further Reading

Dean PDG, Johnson WS and Middle FA (1984) Affinity Chromatography: A Practical Approach. Oxford: IRL Press.

Fassina G and Chaiken IM (1987) Analytical high‐performance affinity chromatography. Advances in Chromatography 27: 247–297.

Hermanson GT, Krishna Mallia A and Smith PK (1992) Immobilized Affinity Ligand Techniques. Orlando: Academic Press.

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How to Cite close
Fassina, Giorgio(Apr 2001) Affinity Chromatography. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0001111]