Abstract

Large volume sample stacking using electrosmotic flow pump (LVSEP) in capillary electrophoresis is a useful tool for on-line concentration of low-abundant protein samples for proteomics research. While the stacking of only small anions with high p K a values has been reported at low pH buffers (pH<4), protein samples (trypsin inhibitor and α-lactalbumin) with high p I values (p I>4) were stacked successfully in our system using LVSEP. Initially, the whole column of the SIL, or LPA-coated capillary was filled with protein samples (∼1 μg/ml), then a high voltage was applied without polarity switching with high pH buffer (pH 8.5). An enhancement factor of more than 100 times was achieved compared to the conventional pressure injection method.

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