Generation of peptide maps by capillary zone electrophoresis in isoelectric iminodiacetic acid

Electrophoresis
A Bossi, P G Righetti

Abstract

Capillary zone electrophoresis in stationary, isoelectric buffers is a novel method for generating peptide maps of protein digests. The buffer system developed is composed of iminodiacetic acid (IDA), whose physico-chemical parameters were found -- by theoretically modeling and experimental verification -- to be: pI 2.23 (at 100 mM concentration), pK1 = 1.73 and pK2 = 2.73 (no attempts were made at measuring the pK of the primary amino group, since such a low pI value would be compatible with any pK value of the basic group, down to as low as pK 5.5). IDA is compatible with most hydro-organic solvents, including trifluoroethanol (TFE), up to at least 40% v/v, typically used for modulating peptide mobility. In naked capillaries, a buffer comprising 50 mM IDA, 10% TFE and 0.5% hydroxyethylcellulose (HEC) allows generation of peptide maps with high resolution, reduced transit times and no interaction of even large peptides with the wall. However, the best background electrolyte was found to be a solution of 50 mM IDA in 0.5% HEC and 6-8 M urea, one of the best solubilizers of proteins and peptides known. In this last electrolyte system, peptide maps of beta-casein digests (known to contain also very large peptides, up to 6000 Da) ...Continue Reading

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Citations

May 27, 1999·Electrophoresis·A V Stoyanov, P G Righetti
May 18, 1999·Journal of Chromatography. a·E StellwagenP G Righetti
Jul 25, 2000·Journal of Chromatography. a·S R Bean, G L Lookhart
Sep 11, 1999·Journal of Biochemical and Biophysical Methods·P G RighettiC Gelfi
Jan 8, 2000·Journal of Biochemical and Biophysical Methods·P G Righetti, C Gelfi
Sep 16, 1999·Journal of Chromatography. a·A BossiP G Righetti
Jul 31, 2014·Organic & Biomolecular Chemistry·Mario MarciaAndreas Hirsch
Feb 2, 2008·Journal of Chromatography. a·Jean-Marc BusnelGabriel Peltre
Dec 29, 2005·Electrophoresis·Jean-Marc BusnelGabriel Peltre
Dec 31, 2005·Journal of Chromatography. a·Michael TilleyKatherine A Tilley
Nov 2, 2014·Journal of Separation Science·Theresa Kristl, Hanno Stutz
Apr 12, 2001·Electrophoresis·P G RighettiL Castelletti
Dec 28, 2017·Critical Reviews in Food Science and Nutrition·Bindu A Sunilkumar, Eden Tareke
Apr 23, 2008·Current Protocols in Protein Science·P G RighettiC Gelfi
Dec 22, 2019·Applied Biochemistry and Biotechnology·Igor Tadeu Lazzarotto BresolinSônia Maria Alves Bueno
Dec 22, 1999·Electrophoresis·V Kasicka

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