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Dye Affinity Chromatography: History
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Contributor: Helena Kang

Immobilized textile dyes, most commonly sulfonated triazine dyes such as Cibacron Blue F3GA, constitute the ligand class that defines dye affinity chromatography. The dye is covalently attached to a hydrophilic chromatographic matrix and presents a planar, charged, and hydrogen-bonding surface that mimics nucleotide cofactors or other binding pockets, so that a subset of proteins, enzymes, and certain polysaccharides is retained by a combination of electrostatic, hydrophobic, and pseudoaffinity contacts [1]. Elution is effected by a competing ligand, by a change of ionic strength or pH, or by additives that weaken the dye–solute contact [2]. The method is therefore a form of affinity chromatography whose selectivity originates in the dye structure rather than in an antibody, lectin, or immobilized metal. Conceptual boundaries exclude ordinary ion-exchange or hydrophobic-interaction chromatography that lacks a dye ligand, and they also exclude free-solution dye precipitation. Essential features are a chemically defined immobilized dye, a matrix that does not itself dominate retention, and a reversible binding equilibrium that can be displaced without destroying the target [3]. Retention of both sulfated polysaccharides and nucleotide-binding enzymes confirms that selectivity follows dye–solute recognition rather than membership in a single analyte class [4].

  • dye affinity chromatography
  • triazine dye
  • Cibacron Blue
  • ligand chromatography

Chromatography in Natural Products • Analytical Chemistry • Chemistry • Physical Sciences

References

  1. Tony Atkinson; Peter M. Hammond; Roy D. Hartwell; Peter Hughes; Michael D. Scawen; Roger F. Sherwood; David A. P. Small; Chris J. Bruton; Michael J. Harvey; Chris R. Lowe; et al. Triazine-dye affinity chromatography. Biochem. Soc. Trans. 1981, 9, 290-293, 10.1042/bst0090290.
  2. Tsutomu Arakawa; Daisuke Ejima; Kouhei Tsumoto; Matsujiro Ishibashi; Masao Tokunaga; Improved performance of column chromatography by arginine: Dye-affinity chromatography. Protein Expr. Purif. 2007, 52, 410-414, 10.1016/j.pep.2006.10.005.
  3. David H. Sharkis; Richard P. Swenson; Purification by cibacron blue F3GA dye affinity chromatography and comparison of NAD(P)H:quinone reductase (E.C.1.6.99.2) from rat liver cytosol and microsomes. Biochem. Biophys. Res. Commun. 1989, 161, 434-441, 10.1016/0006-291x(89)92617-x.
  4. Ahmed Zayed; Kai Muffler; Thomas Hahn; Steffen Rupp; Doris Finkelmeier; Anke Burger-Kentischer; Roland Ulber; Physicochemical and Biological Characterization of Fucoidan from Fucus vesiculosus Purified by Dye Affinity Chromatography. Mar. Drugs 2016, 14, 79, 10.3390/md14040079.
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