Nanoflow LC/IMS-MS and LC/IMS-CID/MS of protein mixtures

Reña A. Sowell, Stormy L. Koeniger, Stephen J. Valentine, Myeong Hee Moon, David E. Clemmer

Research output: Contribution to journalArticlepeer-review

33 Citations (Scopus)

Abstract

A simple ion trap/ion mobility/time-of-flight (TOF) mass spectrometer has been coupled with nanoflow liquid chromatography to examine the feasibility of analyzing mixtures of intact proteins. In this approach proteins are separated using reversed-phase chromatography. As components elute from the column, they are electrosprayed into the gas phase and separated again in a drift tube prior to being dispersed and analyzed in a TOF mass spectrometer. The mobilities of ions through a buffer gas depend upon their collision cross sections and charge states; separation based on these gas-phase parameters provides a new means of simplifying mass spectra and characterizing mixtures. Additionally it is possible to induce dissociation at the exit of the drift tube and examine the fragmentation patterns of specific protein ion charge states and conformations. The approach is demonstrated by examining a simple three-component mixture containing ubiquitin, cytochrome c, and myoglobin and several larger prepared protein mixtures. The potential of this approach for use in proteomic applications is considered.

Original languageEnglish
Pages (from-to)1341-1353
Number of pages13
JournalJournal of the American Society for Mass Spectrometry
Volume15
Issue number9
DOIs
Publication statusPublished - 2004 Sep

Bibliographical note

Funding Information:
This work is supported by a grant from the National Institutes of Health (1R01GM-59145-03). The authors are also grateful for support from the Indiana Genomics Initiative (INGEN), which has funded some instrument development in our group that is related to this work. MHM is supported under the INGEN funds.

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Spectroscopy

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