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Two dimensional mapping of cancer cell extracts by liquid chromatography-capillary electrophoresis with ultraviolet absorbance detection

  1. Author:
    He, Y.
    Yeung, E. S.
    Chan, K. C.
    Issaq, H. J.

  2. Author Address

    Iowa State Univ Sci & Technol, Ames Lab, USDOE, Ames, IA 50011 USA Iowa State Univ Sci & Technol, Ames Lab, USDOE, Ames, IA 50011 USA Iowa State Univ Sci & Technol, Dept Chem, Ames, IA 50011 USA NCI Frederick, SAIC Frederick, Frederick, MD 21702 USA Yeung ES Iowa State Univ Sci & Technol, Ames Lab, USDOE, Ames, IA 50011 USA
    1. Year: 2002
  2. Journal: Journal of Chromatography A
    1. 979
    2. 1-2
    3. Pages: 81-89
  4. Type of Article: Article
  1. Abstract:

    A two-dimensional liquid chromatography/capillary electrophoresis technique was developed for rapid and comprehensive mapping of cell extracts. The cell extracts were first separated by reversed-phase HPLC based on hydrophobicity. Fractions of the effluent from the HPLC system were collected into 96-well microtiter plates and dried under vacuum. The fractions were reconstituted with deionized water, separated by capillary array electrophoresis based on charge-to-size ratio, and detected by UV absorption at 214 nm. Prior to analysis by multiplexed capillary electrophoresis, the reconstituted fractions were concentrated on-column using large volume sample stacking with polarity switching. In this way, high-resolution analysis of even the minor components in the complicated mixture was possible. (C) 2002 Elsevier Science B.V. All rights reserved.

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