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

Abstract

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.
Pages95-103
Number of pages9
ISBN (Print)9780124201194
DOIs
StatePublished - 2014

Publication series

NameMethods in Enzymology
Volume541
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • 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

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  • Cite this

    Duong-Ly, K. C., & Gabelli, S. B. (2014). Using ion exchange chromatography to purify a recombinantly expressed protein. In Laboratory Methods in Enzymology: Protein Part C (pp. 95-103). (Methods in Enzymology; Vol. 541). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-420119-4.00008-2