Dye-ligand affinity adsorbents for enzyme purification
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Affinity chromatography is widely employed in laboratory and large-scale for the purification of biotherapeutics and diagnostics. Some of the most widely used ligands in affinity chromatography have been several reactive chlorotriazine dyes. In particular, immobilized anthraquinone dyes have found a plethora of applications in affinity chromatography because they are inexpensive, are resistant to chemical and biological degradation, are sterilizable and cleanable in situ, and are readily immobilized to generate affinity absorbents which display high binding capacity for a broad spectrum of proteins.
This article provides detailed protocols on the preparation of a dye-ligand affinity adsorbent. Also, detailed protocols for effective application of these media, emphasizing binding and elution conditions are presented.
Index EntriesAffinity chromatography biomimetic dyes chlorotriazine dyes protein purification
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- 4.Labrou, N. E. and Clonis, Y. D. Immobilised synthetic dyes in affinity chromatography. In: Vijayalakshmi, M. A. (ed.): Theory and Practice of Biochromatography. Harwood Academic Publishers, Netherlands, (in press).Google Scholar
- 15.Labrou, N. E. (2000) Dye Affinity Labelling of yeast Alcohol Dehydrogenase. Enzyme Inhibition 15, 487–496.Google Scholar
- 17.Small, D. A., Lowe, C. R., Atkinson, T., and Bruton, C. J. (1982) Affinity labelling of enzymes with triazine dyes. Isolation of a peptide in the catalytic domain of horse liver alcohol dehydrogenase using Procion Blue MX-R as a structural probe. Eur. J. Biochem. 128, 119–123.PubMedCrossRefGoogle Scholar