Using ion exchange chromatography to purify a recombinantly expressed protein

Krisna C. Duong-Ly, Sandra B. Gabelli

Research output: Chapter in Book/Report/Conference proceedingChapter

8 Scopus citations


Ion exchange chromatography (IEX) separates molecules by their surface charge, a property that can vary vastly between different proteins. There are two types of IEX, cation exhange and anion exchange chromatography. The protocol that follows was designed by the authors for anion exchange chromatography of a recombinantly expressed protein having a pI of 4.9 and containing two cysteine residues and one tryptophan residue, using an FPLC system. Prior to anion exchange, the protein had been salted out using ammonium sulfate precipitation and partially purified via hydrophobic interaction chromatography (see Salting out of proteins using ammonium sulfate precipitation and Use and Application of Hydrophobic Interaction Chromatography for Protein Purification). Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.

Original languageEnglish (US)
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationProtein Part C
PublisherAcademic Press Inc.
Number of pages9
ISBN (Print)9780124201194
StatePublished - 2014

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • Carboxymethyl (CM)
  • Diethylaminoethyl (DEAE)
  • FPLC system
  • Ion exchange chromatography (IEX)
  • Quaternary aminoethyl (QAE)
  • Quaternary anion (Q)
  • Sulfopropyl (SP)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


Dive into the research topics of 'Using ion exchange chromatography to purify a recombinantly expressed protein'. Together they form a unique fingerprint.

Cite this