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Ion Exchange Chromatography: History
Please note this is an old version of this entry, which may differ significantly from the current revision.
Contributor: Helena Kang

Reversible electrostatic binding of charged solutes to an oppositely charged immobilized exchanger, followed by elution with a competing ion or a pH change, defines ion-exchange chromatography (IEX). Cation exchangers carry sulfonate or carboxylate groups; anion exchangers carry quaternary ammonium or diethylaminoethyl groups; both may be realized on resins, celluloses, silicas, or thin layers [1]. Retention is a function of the solute’s net charge, the exchanger’s capacity and pKa, and the ionic strength and pH of the buffer [2]. Modern IEX of biopharmaceuticals uses high-resolution particle and monolith formats to resolve charge variants of proteins, which remains the same electrostatic mechanism [3]. Conceptual boundaries exclude reversed-phase and hydrophilic-interaction chromatography, affinity chromatography, and electrophoretic separations that lack a chromatographic exchanger. Essential features are a defined ionogenic ligand, a buffered aqueous mobile phase, and a salt or pH program that displaces bound ions. Urea and related additives may modulate secondary (non-ionic) contacts without changing the primary ion-exchange definition [4].

  • ion-exchange chromatography
  • cation exchanger
  • anion exchanger
  • electrostatic binding

Analytical Chemistry and Chromatography • Spectroscopy • Chemistry • Physical Sciences

References

  1. K. Randerath; E. Randerath; Ion-exchange chromatography of nucleotides on poly-(ethyleneimine)-cellulose thin layers. J. Chromatogr. A 1964, 16, 111-125, 10.1016/s0021-9673(01)82445-6.
  2. Tony E. Hugli; Stanford Moore; Determination of the Tryptophan Content of Proteins by Ion Exchange Chromatography of Alkaline Hydrolysates. J. Biol. Chem. 1972, 247, 2828-2834, 10.1016/s0021-9258(19)45285-x.
  3. Szabolcs Fekete; Alain Beck; Jean-Luc Veuthey; Davy Guillarme; Ion-exchange chromatography for the characterization of biopharmaceuticals. J. Pharm. Biomed. Anal. 2015, 113, 43-55, 10.1016/j.jpba.2015.02.037.
  4. R. V. Tomlinson; G. M. Tener; The Effect of Urea, Formamide, and Glycols on the Secondary Binding Forces in the Ion-Exchange Chromatography of Polynucleotides on DEAE-Cellulose*. Biochem. 1963, 2, 697-702, 10.1021/bi00904a013.
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