LabAutomation 2006 - SLAS

LabAutomation2006
12:00 pm Monday, January 23, 2006 Track 1: Detection & Separation Room: Catalina
Wyndham Palm Springs Hotel
John E. P. Syka
Co-Author(s)
Thermo Electron Co./University of Virginia
Joshua C. Coon, University of Wisconsin
Charlottesville, Virginia
john.syka@thermo.com
Beatrix M. Ueberheide, Rockefeller University
An Ch, University of Virginia
Lewis Y. Geer, National Library of Medicine NIH
Leann M. Hopkins, Dina L. Bai, Donald F. Hunt, University of Virginia
Electron Transfer Dissociation (ETD): Extending the Reach of Mass Spectrometry in
Peptide and Protein Analysis
Electron transfer dissociation (ETD) has rapidly developed into a powerful analytical technique. We have modified a Finnigan LTQ RF 2D
ion trap mass spectrometer by adapting a negative ion chemical ionization source to the back end of the instrument and by modifying the
2D trap electronics to allow charge sign independent ion confinement during ion/ion reactions. Under software control, various CAD, ETD
and proton transfer charge reduction (PTR) events can be incorporated into a basic MS/MS experiment by insertion of the appropriate
ion manipulation steps We have identified reagent ions that provide electron transfer efficiencies exceeding 70%, as well as ion source
compatible PTR reagent anions enabling such experiments to be performed at analytically useful sample levels within the context of online
LC MS/MS analyses. Phosphorylated peptides exhibit facile loss of phosphoric acid in CAD. ETD, like ECD (electron capture dissociation),
yields c/z type fragmentation without loss of CAD labile moieties and generally yields product ions representing most all backbone cleavage
sites. We have demonstrated the utility of ETD in the analysis of phosphopeptides via data-dependent LC MS/MS analysis of a whole yeast
lysate sample. We also have found that data dependent LC MS/MS with ETD/PTR very effective for the analysis of the complex mixtures of
variably post-translationally modified Histone peptides. This ETD/PTR technique has also been applied to small proteins to yield respective
N and C terminal c/z type ion series which can be utilized for direct identification of the precursor via data base search.
3:00 pm Monday, January 23, 2006 Track 1: Detection & Separation Room: Catalina
Wyndham Palm Springs Hotel
Tom Lloyd
Wyeth Research
Collegeville, Pennsylvania
lloydt@wyeth.com
Flexible Automation Tools for Compound Optimization and Pre-Clinical Drug
Metabolism
A variety of automation tools ranging from standalone workstations, to integrated robotics systems, to on-line LC/MS/MS systems will be
discussed. Examples will be presented of how these tools are applied in support of in vivo and in vitro studies at different stages of drug
development. Rapid extraction, incubation and LC/MS/MS method development tools will be discussed including a generic turbulent
flow chromatography assay, multiplexing, parallel chromatography, and versatile on-line analysis systems. Applications will include protein
binding, CYP450 in vitro assays and PK analysis including working with whole blood and homogenized brain tissue.
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