High-performance capillary electrophoresis using open tubes and gels |
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Authors: | A S Cohen A Paulus B L Karger |
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Institution: | (1) Barnett Institute and Department of Chemistry, Northeastern University, 02115 Boston, Massachusetts, USA |
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Abstract: | Summary This paper overviews several aspects of high performance capillary electrophoresis (HPCE), a promising new method of analytical
and micropreparative separation of biochemically important samples. The basic migration equations of electrophoresis are first
presented and the benefit of high fields for rapid analysis and high performance emphasized. Since power is generated with
high voltages, Joule heating results and this heat must be dissipated. The use of capillary columns is shown to be important
in efficient heat removal and in minimizing the temperature differences within the column. The various factors influencing
band broadening are next described, and it is shown how plate counts close to 106 can be achieved. Various results from our laboratory on open tube and gel columns are then presented to illustrate the potential
of this method. Chiral resolution of dansylated amino acids using a chiral metal chelate micelle in open tube HPCE is shown.
With the gel columns, the baseline separation of a 2-chain variant from methionine growth hormone (met-hGH) under non-denaturing
conditions at fields close to 1000 V/cm is presented. Finally, the micropreparative purification of a 20-mer oligonucleotide
using the gel column is described. |
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Keywords: | Capillary electrophoresis Capillary gelelectrophoresis Micropreparative electrophoresis |
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