287438 Isoelectric Focusing without Carrier Ampholytes

Tuesday, October 30, 2012: 8:45 AM
Fayette (Westin )
Frank Jahnke and Penny Ross, Sonata Biosciences, Inc., Auburn, CA

Analytical applications of isoelectric focusing use carrier ampholytes to maintain a linear pH gradient between the anolyte and catholyte.  There are applications where use of carrier ampholytes is troublesome, including subsequent sample analysis by mass spectrometry. 

We report here on capillary isoelectric focusing when carrier ampholytes are removed.  This is motivated partially by presentations at last year's AES meeting, where results were presented and interpreted that differ considerably from ours.  We report on the nature of the pH gradient measured by the change of speciation, ultraviolet absorbance and fluorescence of molecular pH probes.  These results are compared with theory derived from first principles.  Finally we show how single proteins and simple protein mixtures focus in such a carrier-ampholyte-free system using imaging capillary isoelectric focusing.

This text is added to conform with the stated requirement on the minimum word count of an abstract that will be accepted by the American Institute of Chemical Engineers and the American Electrophoresis Society.


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