Micellar electrokinetic chromatography of fluorescently labeled proteins on poly(dimethylsiloxane)-based microchips

Electrophoresis. 2006 Jul;27(14):2933-9. doi: 10.1002/elps.200500795.

Abstract

MEKC of standard proteins was investigated on PDMS microfluidic devices. Standard proteins were labeled with AlexaFluor(R) 488 carboxylic acid tetrafluorophenyl ester and filtered through a size-exclusion column to remove any small peptides and unreacted label. High-efficiency MEKC separations of these standard proteins were performed using a buffer consisting of 10 mM sodium tetraborate, 25 mM SDS, and 20% v/v ACN. A separation of BSA using this buffer in a 3.0 cm long channel generated a peak with a plate height of 0.38 microm in <20 s. Additional fast separations of myoglobin, alpha-lactalbumin, lysozyme, and cytochrome c also yielded peaks with plate heights ranging from 0.54 to 0.72 microm. All proteins migrated with respect to their individual pIs. To improve the separations, we used a PDMS serpentine chip with tapered turns and a separation distance of 25 cm. The number of plates generated increased linearly with increasing separation distance on the extended separation channel chips; however, the resolution reached an asymptotic value after about 7 cm. This limited the peak capacity of the separation technique to 10-12.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Buffers
  • Chromatography, Micellar Electrokinetic Capillary*
  • Dimethylpolysiloxanes / chemistry*
  • Fluorescence
  • Fluorescent Dyes / chemistry
  • Fluorobenzenes / chemistry
  • Humans
  • Microfluidic Analytical Techniques*
  • Proteins / analysis*
  • Proteins / chemistry
  • Sensitivity and Specificity

Substances

  • AlexaFluor 488 carboxylic acid tetrafluorophenyl ester
  • Buffers
  • Dimethylpolysiloxanes
  • Fluorescent Dyes
  • Fluorobenzenes
  • Proteins