Desalting Electroeluted Proteins with Hydrophilic Interaction Chromatography

P. Jeno, P. E. Scherer, U. Manningkrieg, M. Horst

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

We describe a chromatographic procedure for the removal of sodium dodecyl sulfate (SDS) from proteins isolated by electroelution. It involves chromatography of electroeluates on poly(2-hydroxyethyl-aspartamide) silica, a support initially developed for hydrophilic interaction chromatography. The electroeluate, dialyzed against ammonium bicarbonate-SDS buffer, is directly injected onto the column, which is equilibrated in an n-propanol concentration greater than 60%. Bound proteins are eluted with a gradient of decreasing n-propanol. This procedure removes essentially all of the Coomassie blue-related contaminants and separates SDS from the protein. Due to the use of volatile buffer systems, the proteins are recovered in completely salt-free form, which facilitates further protein manipulation. After removal of the organic solvent from the chromatographic desalting step, the recovered proteins are directly amenable to N-terminal protein sequencing and, after evaporation of the organic phase, to enzymatic digestions and subsequent separation of fragments by reverse-phase HPLC.

Original languageEnglish (US)
Pages (from-to)292-298
Number of pages7
JournalAnalytical Biochemistry
Volume215
Issue number2
DOIs
StatePublished - Dec 1993

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Salt removal
Chromatography
Hydrophobic and Hydrophilic Interactions
Sodium Dodecyl Sulfate
1-Propanol
Proteins
Buffers
Protein Sequence Analysis
Silicon Dioxide
Digestion
Organic solvents
Salts
High Pressure Liquid Chromatography
Evaporation
Impurities

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

Desalting Electroeluted Proteins with Hydrophilic Interaction Chromatography. / Jeno, P.; Scherer, P. E.; Manningkrieg, U.; Horst, M.

In: Analytical Biochemistry, Vol. 215, No. 2, 12.1993, p. 292-298.

Research output: Contribution to journalArticle

Jeno, P. ; Scherer, P. E. ; Manningkrieg, U. ; Horst, M. / Desalting Electroeluted Proteins with Hydrophilic Interaction Chromatography. In: Analytical Biochemistry. 1993 ; Vol. 215, No. 2. pp. 292-298.
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