Involvement

This list of possible projects is just a few examples of what could be done. It is our hope that our potential relationship with your laboratory can be versatile and flexible enough to provide for your dynamic needs.

Biomarker Identifications

One area where the Kelleher Lab has already proved useful to many collaborators is in the area of biomarker MS/MS and identification. These markers come from serum samples and range between 5 and 15 kDa in size. Through the variety of fragmentation techniques and database analysis tools the Center has at its disposal, purified markers can be identified in a relatively high throughput format. This could also include cases where MS/MS data has already been collected, but MASCOT or SEQUEST has been unable to retrieve a positive identification.

Recombinant Proteins

It is helpful for quality control purposes to verify the mass, sequence, and post-translational state of a recombinant protein after expression in bacteria or eukaryotic cell lines. The normal assumption is that nothing has caused the gene/transcript/protein pathway to change through the transformation or transfection, but that is not always the case. Purified recombinant proteins below 70 kDa can be run on the MS instrumentation available to the Center. A detailed standardized report for each protein can be generated, outlining the verification of protein primary structure or the structural deviations of the protein, as appropriate.

Phospho-peptide Site Mapping

Another area of interest in the field is the analysis of phospho-proteins and mapping of phosphorylation sites. Using large-peptide digestion methods followed by electron capture dissociation (ECD) on the Q-FTMS instrument, phospho-peptides up to ~30 kDa can be fully characterized. Site-specific information would be related back to the corporate partner in the form of a detailed standardized report.

Antibody Characterization SOP development

A more involved project would be to develop a standard operating procedure for the characterization of antibodies of importance to the corporate partner. Such a project would require development in the isolation of light chain vs. heavy chain antibodies, possible digestion strategies for hybrid top down/bottom up characterization (i.e., "middle down" protein characterization), and LC-MS/MS optimization of the ensuing large peptides. The project may also involve use of the multiple fractionation steps (chromatofocusing, for example) to obtain optimal fractionation of proteins/peptides prior to FTMS analysis.

Other Projects

The Center is always open to discussing other potential projects that are of interest to our partners. Please contact us and put us to work for you!

Contact Information

If you are interested in the capabilities offered by the CTDP, please contact Dr. Andrew Forbes via phone (217-333-6607) or e-mail (ajforbes@uiuc.edu).