Simple luminescence detectors using a light-emitting diode or a Xe lamp, optical fiber and charge-coupled device, or photomultiplier for determining proteins in capillary electrophoresis: a critical comparison

Anal Biochem. 2007 Jun 1;365(1):82-90. doi: 10.1016/j.ab.2007.02.021. Epub 2007 Feb 23.

Abstract

The performance of two homemade fluorescence-induced capillary electrophoresis detectors, one based on light-emitting diode (LED) as the excitation source and a charge-coupled device (CCD) photodetector and the other based on a commercial luminescence spectrometer (Xe lamp) as the excitation source and a photomultiplier tube as a detector, were compared for the determination of fluorescent proteins R-phycoerythrin and B-phycoerythrin. Both devices use commercially available, reasonably priced optical components that can be used by nonexperts. After fine optimization of several optical and separation parameters in both devices, a zone capillary electrophoresis methodology was achieved with 50mM borate buffer (pH 8.4) and 10mM phytic acid for the determination of two phycobiliproteins. Detection limits of 0.50 and 0.64microg/ml for R-phycoerythrin and B-phycoerythrin, respectively, were achieved by using the LED-induced fluorescence capillary electrophoresis (LED-IF-CE) system, and corresponding detection limits of 2.73 and 2.16microg/ml were achieved by using the Xe lamp-IF-CE system. Analytical performance and other parameters, such as cost and potential to miniaturization, are compared for both devices.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrophoresis, Capillary / instrumentation*
  • Electrophoresis, Capillary / methods*
  • Equipment Design / instrumentation*
  • Fiber Optic Technology*
  • Luminescence*
  • Optical Fibers
  • Phycobiliproteins / analysis
  • Phycobiliproteins / chemistry
  • Phycoerythrin / analysis*
  • Phycoerythrin / chemistry
  • Proteins / analysis*
  • Sensitivity and Specificity
  • Xenon

Substances

  • Phycobiliproteins
  • Proteins
  • Phycoerythrin
  • Xenon