Skip NavigationSkip to Content
Return Return to Search Results

Development of a two-dimensional protein-peptide separation protocol for comprehensive proteome measurements

  1. Author:
    Janini, G. M.
    Conrads, T. P.
    Veenstra, T. D.
    Issaq, H. J.

  2. Author Address

    NCI, Analyt Chem Lab, Separat Technol Grp, SAIC Frederick Inc, POB B, Frederick, MD 21702 USA NCI, Analyt Chem Lab, Separat Technol Grp, SAIC Frederick Inc, Frederick, MD 21702 USA Issaq HJ NCI, Analyt Chem Lab, Separat Technol Grp, SAIC Frederick Inc, POB B, Frederick, MD 21702 USA
    1. Year: 2003
  2. Journal: Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
    1. 787
    2. 1
    3. Pages: 43-51
  4. Type of Article: Article
  1. Abstract:

    We have developed an effective two-dimensional fractionation protocol of complex proteome mixtures that extends the ability to conduct more comprehensive proteome measurements. A sample containing intact proteins extracted from Saccharomyces cerevisiae was fractionated by liquid phase isoelectric focusing, followed by tryptic digestion and solid-phase extraction (SPE) clean-up and reversed-phase liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS-MS) of the resultant peptides. The clean-up step is designed to desalt the fractions and rid them of urea and ampholytes prior to analysis by LC-MS-MS. Fifty milligrams of protein were separated into 20 fractions by liquid-phase isoelectric focusing, spanning a pH range of 3-10. The effectiveness of the removal of ampholytes was monitored by capillary zone electrophoresis and LC-MS-MS. The ability to analyze all of the 20 fractions without any noticeable decrease in the separation efficiency demonstrates the overall effectiveness of the SPE clean-up step. The results show that the separation strategy is effective for high throughput characterization of proteins from complex proteomic mixtures. (C) 2002 Elsevier Science B.V. All rights reserved.

    See More

  1. Keywords:

External Sources

  1. No sources found.

Library Notes

  1. No notes added.
NCI at Frederick

You are leaving a government website.

This external link provides additional information that is consistent with the intended purpose of this site. The government cannot attest to the accuracy of a non-federal site.

Linking to a non-federal site does not constitute an endorsement by this institution or any of its employees of the sponsors or the information and products presented on the site. You will be subject to the destination site's privacy policy when you follow the link.

ContinueCancel