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Updated: Jul 3, 2026

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry
Published on: February 7, 2022
Protein chromatography in neat organic solvents
1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Abstract:
Pure formamide and ethylene glycol are used instead of water as processing media for protein chromatography. A number of common proteins are freely soluble in these solvents and most do not undergo irreversible inactivation in them. Batch adsorption studies reveal that proteins readily adsorbed to various ion-exchangers in formamide and ethylene glycol and subsequently can be completely desorbed by adding inorganic salts (LiCl and NH(4)NO(3)) to the solvents. The idea of protein separations in formamide and ethylene glycol is illustrated by column chromatography and preparative separation of mixtures of (i) oxidized A and B chains of insulin and (ii) lysozyme and ribonuclease on the anion-exchanger triethylaminoethycellulose and the cation-exchanger phosphocellulose, respectively.
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