Peptide and Protein Characterization with Mass Spectrometry
Who Should Attend
Protein
chemists who are new to mass spectrometry or are considering
LC/MS or MALDI will benefit from this introduction to the technique.
Experienced MS analysts who find themselves confronting protein
analysis problems will also benefit from the practical advice
and applications overview. All attendees will come away with
a basic knowledge of state-of-the-art approaches to protein analysis
by MS and will find the well-organized course manual a useful
tool to have in the lab as well as an invaluable source of references.
How You'll Benefit from This Course:
- Overiew
of mass spectrometry fundamentals plus the operational specifics
of mass analyzers and LC/MS and MALDI sample introduction systems
employed for protein analysis. Discussions and examples also
include the latest micro and nano-scale systems.
- Practical
advice on development of HPLC methods compatible with LC/MS—sample
preparation, solvent systems, columns and sample considerations
which minimize ion suppression effects and interferences in the
mass spectrum. The effects of different sample matrices in MALDI
will also be discussed.
- Interpretation
of protein mass spectra including sequencing by MS/MS, calculation
of molecular weight from multiple-charge ion electrospray spectra,
and other leading edge applications.
- Presentation
of various laboratory techniques combining digestion, hydrolysis
and derivitization with mass spectrometry to elucidate sequences,
cysteine status and post-translational modifications.
- Data
system applications and Internet resources for analysis of protein
mass spectra and links to databases for determination of unknown
peptides and proteins.
Course Topics:
- Introduction/Review
of Protein Chemistry—amino acids; peptide chemistry and
mass determination; protein structure: primary, secondary, tertiary,
quaternary; transcription, translation, post-translational modifications.
- Mass
Spectrometry Fundamentals—atoms, molecules, ions; the mass
spectrum; isotopes and their importance in MS; fragmentation
and interpretation in MS.
- Mass
Spectrometric Analyzers Used for Protein Characterization—transmission
quadrupole; quadrupole ion trap; time-of-flight; magnetic sector;
detectors and vacuum systems.
- Ionization
and Inlet Techniques Used For Protein Characterization—fast
atom bombardment (FAB); matrix assisted laser desorption ionization
(MALDI); electrospray (ES); microelectrospray, nanoelectrospray
and micro-chromatographic techniques.
- Applications
Overview—molecular weight determination; mass mapping; peptide
sequencing; cysteine status determination; investigation of non-covalent
interactions.
- Optimizing
Mass Spectrometric Results for Peptides and Proteins—MALDI
matrix considerations; electrospray solvent system considerations;
sample preparation procedures.
- Protein
data analysis and database tools—integrated MS data acquisition
and analysis systems; off-line software tools; internet software
tools.
The Instructors:
Frederick E. Klink, STM: Course Director
J. Throck Watson, Michigan State University
Here's What Past Participants Have to Say About This Course:
"This course is a good primer for those looking into the
field." —Michael Evans, Senior Scientist, Cargill Company
©Copyright
2003 Frederick E. Klink and the American Chemical Society.
All rights reserved. |