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Free flow cell electrophoresis using zwitterionic buffer.

L S Rodkey1

  • 1Department of Pathology, University of Texas Medical School, Houston 77225-0708.

Applied and Theoretical Electrophoresis : the Official Journal of the International Electrophoresis Society
|January 1, 1990
PubMed
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A novel zwitterionic buffer demonstrates superior stability and buffering capacity for cell electrophoresis, improving cell separation and maintaining viability. This advancement offers better resolution for fixed cells and reproducible mobility differences in hybridoma cells.

Area of Science:

  • Biophysical Chemistry
  • Cell Biology
  • Biotechnology

Background:

  • Standard buffers used in cell electrophoresis exhibit instability in electrical fields and poor buffering at operational pH.
  • Existing methods lack optimal conditions for precise cell separation and viability maintenance during electrophoresis.

Purpose of the Study:

  • To evaluate a zwitterionic buffer's stability and buffering capacity for cell electrophoresis.
  • To compare the zwitterionic buffer's performance against standard buffers for cell separation.
  • To assess the impact of the zwitterionic buffer on viable hybridoma cell viability and electrophoretic mobility.

Main Methods:

  • Utilized the McDonnell-Douglas Continuous Flow Electrophoresis System for buffer analysis.
  • Conducted stability tests in electrical fields and titration studies for buffering capacity.

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  • Performed comparative cell separation studies with formalin-fixed red blood cells and viable hybridoma cells.

    • Main Results:

    • Standard buffers showed inherent instability and poor buffering at electrophoresis pH.
    • The zwitterionic buffer exhibited excellent stability and buffering capacity.
    • Improved resolution of fixed red blood cells and maintained hybridoma cell viability comparable to Hank's Balanced Salt Solution.
    • Reproducible differences in electrophoretic mobility were observed for hybridoma cells at different growth stages.

    Conclusions:

    • The developed zwitterionic buffer is stable and effective for cell electrophoresis.
    • It offers superior cell separation resolution and supports cell viability.
    • This buffer system provides a reliable tool for analyzing cell electrophoretic properties, including hybridoma cells.