Exploring human plasma proteome strategies: High efficiency in-solution digestion protocol for multi-dimensional protein identification technology

Qinhua Cindy Ru*, Luwang Andy Zhu, Richard A. Katenhusen, Jordan Silberman, Henry Brzeski, Michael Liebman, Craig D. Shriver

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Multi-dimensional protein identification technology (MudPIT) is becoming a prevalent proteomic approach due to its high-throughput separations and accurate mass detection. Prior to MudPIT analysis, complicated samples required in-solution digestion. Unlike in-gel digestion, in which enzymes work on just a few proteins, in-solution digestion involves simultaneous digestion of hundreds or thousands of proteins. In-solution digestion protocols must therefore be very efficient. Few investigations have evaluated the efficiency of in-solution digestion protocols. The present research compared three such protocols. Results suggest that a protocol utilizing trifluoroethanol (TFE) as denaturant is most efficient.

Original languageEnglish
Pages (from-to)175-191
Number of pages17
JournalJournal of Chromatography A
Volume1111
Issue number2
DOIs
StatePublished - 14 Apr 2006
Externally publishedYes

Keywords

  • Clinical proteomics
  • Human plasma
  • Human plasma proteome
  • In-solution digestion
  • Multi-dimensional protein identification technology

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