Mass Spectrometry Laboratory
The BGPF Mass Spectrometry laboratory provides state-of-the-art molecular analysis of biomolecules to UPCI and University of Pittsburgh researchers. Mass spectrometry offers flexible, sensitive and rapid analysis of a wide variety of peptides, proteins, and lipids important to translational research. The lab offers several ionization methods, depending upon the nature of the sample to be analyzed including: electron ionization, fast atom bombardment, and electrospray ionization. Technological advances available at the facility include an upgraded electrospray source to provide nanospray capabilities in the analysis of proteins and peptides.
The facility offers a variety of related services, including mass measurement, assessment of chemical modifications, daughter ion scans (for sequence/structural analysis), neutral loss scans, and selected ion monitoring. The lab can also process larger peptides and proteins for analysis using in-gel digestions of Coomassie or silver-stained gels, solution digests, and digests from polyvinylidene fluoride (PVDF) membranes.
Another technology performed by the laboratory is protein mass
profile fingerprinting, in which a set of peptide mass ions
characteristic of a particular protein are obtained from the protein's
cleavage products. Databases can then be searched to fit the
mass profile fingerprint with known proteins.
The Mass Spectrometry Facility has developed an online HPLC microcapillary system to interface with the mass spectrometer (HPLC-MS). This type of technology extends the sensitivity of the instrument by allowing online concentration of the sample, thereby increasing the signal-to-noise ratio. The microcapillary HPLC-MS application is currently being used for analysis of low-level amounts of peptides for sequence analysis. In addition to the online microcapillary HPLC-MS with nanoflow capabilities, the facility also offers a nanovial option, whereby small amounts of sample (1-5 ml) can be analyzed.
The Mass Spectrometry lab also performs phosphopeptide analysis, and has developed a mass spectrometric method whereby lipids can be analyzed by direct infusion into the mass spectrometer. Quantitation can also be performed in these analyses, which are relevant to studies of apoptosis, liposome technology, and lipid-signaling molecules. The facility has extended its analysis of lipids by mass spectrometry to include subcellular organelle fractions, like mitochondria.