Protein separation and enrichment by counter-current chromatography using reverse micelle solvent systems

Ching Wei Shen, Tiing Yu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

A protein mixture consisting of myoglobin, cytochrome c, and lysozyme was separated by high-speed counter-current chromatography using a two-phase aqueous/reverse micelle-containing organic solvent system. About 50% stationary phase retention ratio was obtained in most chromatographic experiments. Separations were manipulated mainly by pH gradients that controlled the electrostatic interactions between the protein molecules and reverse micelles. Separations were further improved by incorporating an ionic strength gradient along with the pH gradient. Control of ionic strength in the aqueous solution helped fine-tune protein partitioning between the stationary and mobile phases. Although non-specific protein interactions affected baseline resolution, recovery of cytochrome c and lysozyme reached 90% and 82%. Furthermore, concentration or enrichment of these two proteins was achieved from a large-volume sample load. This technique can potentially be employed in the recovery and enrichment of proteins from large-volume aqueous solutions.

Original languageEnglish
Pages (from-to)164-168
Number of pages5
JournalJournal of Chromatography A
Volume1151
Issue number1-2
DOIs
StatePublished - 1 Jun 2007

Keywords

  • Counter-current chromatography
  • Ionic strength gradient
  • pH gradient
  • Protein enrichment
  • Protein separation
  • Reverse micelle

Fingerprint Dive into the research topics of 'Protein separation and enrichment by counter-current chromatography using reverse micelle solvent systems'. Together they form a unique fingerprint.

Cite this