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Overview of Barnett Institute Research Programs
The research groups of Profs. Karger and Hancock are internationally recognized for their contributions to proteomic analysis; the new professors Engen and Zhou complement this expertise with new methods of targeted protein analysis using HX-exchange and biochemical assays. Work in varied collaborations has been fruitful in optimizing all steps of the proteomic workflow. including significant advances in the software for analyzing LC-MS data sets. In the key area of LC-MS analysis, the use of ultra-narrow bore monolithic capillary columns and porous-layer open tubular (PLOT) columns with nanospray ESI has dramatically inproved sensitivity and resolution. Dr. Hancock's Multiple Lectin Affinity Chromatography can select glycoproteins, a highly promising class of serum proteins for biomarker discovery. Biomarkers for cervical cancer have been discovered in collaboration with Cytyc Corp; collaborations are active in many other cancers. Extended Range Proteomic Analysis (ERPA) takes more full advantage of the capabilities of the new hybrid FTMS-linear ion trap mass spectrometers in comprehensive characterization of a targeted protein. Comprehensive protein analysis has revealed significant differences between bona-fide and generic protein therapeutics The NanoSplitter developed in Paul Vouros' group provides the advantages of nanoelectrospray mass spectrometry with LC methods developed on traditional 4 mm columns. 100-improvement in S/N while permitting 99% of the eluant to be directed to fraction collection or other methods of detection, such as UV, radiometric detection or Coulometric Arrays. Micro-NMR methods, have reduced the sample mass to obtain interpretable NMR spectra down to the mid-nanogram level. When used to load LC-MS fraction collected using the NanoSpltter, this system provides an exquisitely sensitive and practical LC-MS-NMR platform. Differential Ion Mobility (DMS) has proven invaluable as a front-end to MS, to disrupt or filter cluster ions. An LC-MS method for profiling 15 conjugated lipid hormones in a single run was developed in Vouros' group, using single reaction monitoring among other innovations, in a collaborative program with the Center for Drug Discovery. The Environmental Cancer Center >Drs Giese and Vouros have been developing better assays for DNA adducts including a novel mass tag with high sensitivity, a study on exposure of hospital workers to the sterilization gas ethylene oxide, and CEC and CE assays for biomarkers of carcingens from grilled meats or cigarette smoke. A new program by Dr. Vouros in collaborations with Helmut Zarbl of the Fred Hutchinson cancer center is investigating the relationship between DNA adduct formation and toxicogenomic processes. Chemical Biology
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