MS&D: Biological Separations and Mass Spectrometry
The 9.4-Tesla Fourier transform ion cyclotron resonance mass spectrometer in the High-Performance Mass Spectrometry Facility is a high-throughput proteomics production prototype and received an R&D 100 Award in 2003.
The Biological Separations and Mass Spectrometry group focuses on development of high-resolution separations and mass spectrometry and their applications in biological research. Our current research emphasis is in the area of proteomics – the study of the entire complement of proteins expressed in a cell under a specific set of conditions at a specific time.
The new technologies exploit nano-scale ultra-high pressure capillary liquid chromatography (LC) separations combined with high accuracy mass measurements using Fourier transform ion cyclotron resonance (FTICR) mass spectrometry to make comprehensive, quantitative, and high-throughput proteome measurements. Our approach involves identifying peptide markers for proteins that can be used for subsequent high-throughput mass spectrometric measurements (i.e., without the need for tandem mass spectrometry) and makes use of very high accuracy mass measurements combined with information from the distinctive separation properties of peptides . The use of stable isotope labels or relative peak intensities of these peptide markers also provides the basis for quantitative measurements. Additional new developments have enabled us to significantly extend the dynamic range of measurements to approximately six orders of magnitude and to investigate very small cell populations, and even single cells.
Technology development proceeds in concert with its applications to a number of biological systems. Initial applications have focused on microbial systems, including Deinococcus radiodurans, Shewanella oneidensis, Rhodobacter sphaerodies and Yersina pesti, and have provided a range of new insights into the proteins actually expressed by these organisms under various conditions. Anaerobic microbial metabolism is of direct relevance to the DOE missions in environmental stewardship (contaminant bioremediation, microbial impacts on global warming through production and sequestration of methane and carbon dioxide), clean and secure energy (methane and H2 from wastes as alternative energy sources), and basic science (cycling of carbon, nitrogen, metals, and radionuclides).
More recent work has centered on extending application of these proteomics technologies to mammalian systems, which pose additional challenges due to their much greater complexity. One early focus has been the human blood plasma proteome due to its broad biomedical applications. Plasma proteome measurements potentially can provide the basis for discovery of protein biomarkers or signatures for virtually every disease state. In turn, such markers can enable much earlier and more effective detection of diseases, and thus lead to implementation of more effective therapies. For example, significant efforts at a number of laboratories are being made to identify distinctive protein biomarkers for early detection of cancers, when treatment is known to be much more effective.
Contact: Richard Smith, Technical Leader