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Ultra-fast two-dimensional microchip electrophoresis using SDS μ-CGE and microemulsion electrokinetic chromatography for protein separations

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Abstract

A poly(methyl methacrylate) microfluidic chip was used to perform a two-dimensional (2-D) separation of a complex protein mixture in short development times. The separation was performed by combining sodium dodecyl sulfate micro-capillary gel electrophoresis (SDS μ-CGE) with microemulsion electrokinetic chromatography (μ-MEEKC), which were used for the first and second dimensions, respectively. Fluorescently labeled Escherichia coli cytosolic proteins were profiled by this 2-D approach with the results compared to a similar 2-D separation using SDS μ-CGE × μ-MEKC (micelle electrokinetic chromatography). The relatively short column lengths (effective length = 10 mm) for both dimensions were used to achieve separations requiring only 220 s of development time. High spot production rates (131 ± 11 spots min−1) and reasonable peak capacities (481 ± 18) were generated despite the fact that short columns were used. In addition, the use of μ-MEEKC in the second dimension was found to produce higher peak capacities compared to μ-MEKC (481 ± 18 for μ-MEEKC and 332 ± 17 for μ-MEKC) due to the higher plate numbers associated with μ-MEEKC.

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Acknowledgment

The authors would like to express their gratitude for financial support of this work by the Louisiana Board of Regents and the National Science Foundation (EPS-0346411). The authors would also like to thank the World Class University (WCU) program for partial financial support of this work.

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Author notes

  1. John K. Osiri

    Present address: Department of Chemical Engineering, Washington State University, Pullman, WA, 99164, USA

  2. Hamed Shadpour

    Present address: Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599, USA

Authors and Affiliations

  1. Department of Chemistry, Louisiana State University, Baton Rouge, LA, 70803, USA

    John K. Osiri, Hamed Shadpour & Steven A. Soper

  2. Center for BioModular Multi-Scale Systems, Louisiana State University, Baton Rouge, LA, 70803, USA

    John K. Osiri, Hamed Shadpour & Steven A. Soper

  3. Department of Mechanical Engineering, Louisiana State University, Baton Rouge, LA, 70803, USA

    Steven A. Soper

  4. Nano-Biotechnology and Chemical Engineering, Ulsan National Institute of Science and Technology, 194 Banyeon-ri Ulju-gun Eonyang-eup, Ulsan, 689-805, South Korea

    Steven A. Soper

Authors
  1. John K. Osiri
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  2. Hamed Shadpour
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  3. Steven A. Soper
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Corresponding author

Correspondence to Steven A. Soper.

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Osiri, J.K., Shadpour, H. & Soper, S.A. Ultra-fast two-dimensional microchip electrophoresis using SDS μ-CGE and microemulsion electrokinetic chromatography for protein separations. Anal Bioanal Chem 398, 489–498 (2010). https://doi.org/10.1007/s00216-010-3914-2

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  • DOI: https://doi.org/10.1007/s00216-010-3914-2

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