omation mbers - Society for Laboratory Automation and Screening

10:30 am Thursday, February 5 Proteomics – Technology 3 Room B1
Gary Schultz
Advion BioSciences, Inc.
30 Brown Road
Ithaca, New York 14850
gschultz@advion.com
Sample Preparation Tips for Sample Enrichment and Direct Elution Nanoelectrospray
Mass Spectrometry Analysis For Enhanced Sensitivity For Protein Characterization
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Co-Author(s)
Geoffrey Rule
Sheng Zhang
Colleen K. Van Pelt
Amie Prince
The sample diversity of proteomics requires analytical techniques that can provide rapid and sensitive
characterization of complex biological systems. Nanoelectrospray mass spectrometry is one technique essential
to every researcher working in this field due to its low sample consumption and sensitivity. NanoESI/MS provides
long spray times useful for performing MS/MS including neutral loss and precursor ion scans. Combining
NanoESI/MS with sample preparation can further improve sensitivity when sample enrichment or concentration
is utilized. The NanoMate 100 with ESI Chip is an automated nanoelectrospray system developed to improve
the efficiency and quality of NanoESI/MS. In operation, this system establishes an electric field localized at
the exit of each microfabricated nozzle resulting in a stable, robust spray. That combined with the repeatable
structure of the nozzles provides an automated platform for analysis of microliter sample volumes. Advantages
of the system include low sample consumption, conservation of sample not consumed in the analysis, one-time
spray optimization, enhanced spray stability, and no carryover. Sample preparation tips are routinely used for
desalting and cleanup of proteins. The NanoMate functionality has been enhanced to enable the direct elution-
NanoESI/MS of samples from sorbent-filled pipette tips. Low levels of peptides can be concentrated and enriched.
Analytes are eluted in sub-microliter volumes at 100 nL/min delivering high analyte concentrations to the mass
spectrometer. Applications of on-line elution will be demonstrated for protein digest analysis and identification of
phosphopeptides and glycopeptides.
11:00 am Thursday, February 5 Proteomics – Technology 3 Room B1
Neil Kelleher
University of Illinois
600 S. Mathews Avenue
Urbana, Illinois 61801
kelleher@scs.uiuc.edu
Progress in Automating Top Down Proteomics
Co-Author(s)
Steven Patrie, Dana Robinson,
Yi Du, Lihua Jiang,
Michael Roth
An emergent “Top Down” approach to analysis of intact proteins will be described. Efforts in our laboratory
combine informatics with a Quadrupole/Fourier-Transform hybrid mass spectrometer (Q-FTMS) to enable efficient
characterization of biological events that change the mass of protein molecules from that predicted by an
annotated genome sequence. A platform dedicated to Top Down is under development and uses a size-dependent
proteome fractionation up front, followed by a Q-FTMS engine for data acquisition, and finally a custom database
and software suite called “ProSight PTM” for streamlined protein identification and characterization (Anal. Chem.,
2003, 75, 4081-4086). Protein examples from yeast and human cells will be described, including characterization
of histone modifications using a new database strategy termed “prescriptive annotation”.