Capillary zone electrophoresis of oligonucleotides in isoelectric buffers and against a stationary pH gradient

Electrophoresis
A V StoyanovP G Righetti

Abstract

Capillary zone electrophoresis of oligonucleotides in a background electrolyte of two different types of stationary buffers is proposed: single, isoelectric amphoteres and focused carrier ampholytes. In the first case, two zwitterionic molecules are evaluated: lysine and histidine. Although the former has a five times higher buffering power (beta) at the pI (9.74) than the latter (pI 7.47), due to the favorable delta pK value (1.6 vs. 3) and thus should be the preferred species, a new parameter for evaluating the performance of isoelectric buffers is proposed: the beta/lambda ratio, i.e., the ratio between the buffering power and its conductivity. Ideal buffers are those with the highest beta/lambda ratio, since this allows delivering very high voltage gradients with minimal Joule effects. Since the pI of Lys is situated in a pH region (9.74) where bulk water begins to conduct, whereas His has a pI close to neutrality, the beta/lambda ratio is more favorable for His than for Lys. In the second case (zone electrophoresis of oligonucleotides against a preformed pH gradient), it is shown that migration against a pH 6.5-10 Pharmalyte carrier ampholyte pH gradient offers a unique analyte resolution. This is possibly due to two effec...Continue Reading

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Citations

Feb 12, 1998·Electrophoresis·P G Righetti, C Gelfi
May 18, 1999·Journal of Chromatography. a·N C StellwagenP G Righetti
Jun 17, 1998·Forensic Science International : Synergy·P G Righetti, C Gelfi
Feb 16, 2005·Annual Review of Physiology·Michael Gekle
Jan 8, 2000·Journal of Biochemical and Biophysical Methods·P G Righetti, C Gelfi

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