Current trends in capillary isoelectric focusing of proteins

Pier Giorgio Righetti, Cecilia Gelfi, Monica Conti

Research output: Contribution to journalArticlepeer-review


Isoelectric focusing (IEF) in thin capillaries is reviewed here. After an introduction on the genesis and chemistry of the carrier ampholyte buffers, different approaches to IEF are discussed and evaluated. The classical approach consists on IEF under conditions of suppressed electroosmotic (EOF) flow, usually obtained by covalently bonding hydrophilic polymers to the inner capillary wall. The other approach consists of IEF in dynamically (and partially) coated capillaries, so as to allow a reduced EOF flow to coexist with the IEF process, so that focusing and transport of the train of stacked bands occurs simultaneously. The various experimental parameters: focusing, elution and detection steps, pI measurements, as well as typical drawbacks, such as isoelectric precipitation are evaluated. The review ends with some examples of analytical separations, at the moment mostly limited to focusing of native hemoglobins (normal and point mutants). These separations are compared with those obtained by slab-gel IEF and in immobilized pH gradients.

Original languageEnglish
Pages (from-to)91-104
Number of pages14
JournalJournal of Chromatography B: Biomedical Applications
Issue number1-2
Publication statusPublished - Oct 10 1997


  • Proteins
  • Reviews

ASJC Scopus subject areas

  • Chemistry(all)


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