Abstract
We are now in the era of the proteomics where the interest is in global analysis of many, if not all, proteins in cells, tissues, etc.  At any given time point, thousands of proteins exist in a given cell with 10⁶  levels difference of concentration.  This dynamic range in concentration reaches as high as 10¹²  in plasma.  Furthermore, digestion of individual proteins yield, on average, 40 or more peptides.  As a consequence, proteome analysis of whole cell lysates, organelles or body fluids such as plasma yield enormously complex challenges.  While mass spectrometric instrumentation continues to improve in sensitivity, resolution, mass accuracy and structure elucidation, the front-end sample processing steps prior to the mass spectrometer remain critical.  Multidimensional separation is a requirement in order to simplify mixtures eluting into the mass spectrometer.  In addition, in many cases, very high resolution LC is needed as a final step into the mass spectrometer.  In this talk, we shall overview some of the strategies possible for separation prior to mass spectrometry.  We will present results using monolithic columns, which generate high peak efficiency and at the same time allow use of narrow bore columns 25um i.d. or less for high sensitivity ESI.  We are using such columns for biological samples in which the sample amount is limited, e.g. laser capture microdissection of tumor cells.  In a second example, we are employing high resolution LC coupled to (off-line) MALDI-MS for analysis.  We will demonstrate several advantages of this approach such as the ability to minimize chemical noise as a result of the chromatographic separation and the ability to optimize the MS/MS analysis for specific regions in the chromatogram of interest.  Finally, we will illustrate the use of these and other approaches for quantitative proteomics and biomarker discovery for diagnosis.

Background
Barry Karger received his B.S. in Chemistry from MIT in 1960 and his Ph.D. in Analytical Chemistry from Cornell University in 1963. He then was appointed an Assistant Professor of Chemistry at Northeastern University in Boston, MA, attaining Full Professorship in 1972. In 1973, he co-founded what is now the Barnett Institute of Chemical and Biological Analysis. The Institute, a leading bioanalytical research center, recently celebrated its 30th anniversary. Professor Karger has been the Director of the Institute since its inception. In 1985, he was appointed the first holder of the James L. Waters Chair in Analytical Chemistry.

Dr. Karger’s recent research interests have focused on bioanalysis, including high performance DNA separation and analysis for DNA sequencing, and new separations – ESI and MALDI mass spectrometric methods for proteome analysis and new microscale LC columns for high mass sensitivity, LC/MS. He has been an active researcher with more than 275 publications in the field of separation science, with particular emphasis in liquid chromatography and capillary electrophoresis and most recently, genomics and proteomics. He is a member of the editorial boards of numerous journals in his fields of expertise. He is the holder of 28 patents in these fields, a number of which have been commercialized. With Lloyd Snyder and Csaba Horváth, he is the co-author of An Introduction to Separation Science, a leading graduate textbook. He has trained more than 150 students at the Ph.D. and postdoctoral levels.

In 1989, Barry Karger was encouraged by scientific colleagues to organize the first Symposium on High Performance Capillary Electrophoresis (HPCE) in Boston. The meeting was highly successful and sparked interest in establishing the HPCE meeting as an annual international symposium. Today, this symposium is the major international conference devoted to HPCE and other microscale separation techniques.  Barry, along with a team of Permanent Scientific Committee members, orchestrated the next decade of HPCE meetings, and in 1998 handed over the reins of leadership of this symposium to CaSSS.

Dr. Karger has received numerous honors, including the 2002 Halasz Medal of the Hungarian Chemical Society for Chromatography and Separation Science, the ACS IBC Advanced Technologies and Millipore Award in Separations Science and Technology (1998), ACS Fisher Award in Analytical Chemistry (1990), ACS Supelco Award in Chromatography (1982), EAS Award for Outstanding Achievements in the Field of Separations Science (1998), A.J.P. Martin Medal from the Chromatography Society of the United Kingdom (1991) and the Steven Dal Nogare Memorial Award for Chromatography (1975).
 

Meeting details
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The San Francisco Bay Area Mass Spectrometry discussion group was formed in 1980 to provide a regular gathering for people interested in mass spectrometry and allied topics. BAMS currently has a membership of about 280 individual and 20 corporate members, and meets 8-10 times per year for a midweek dinner and lecture.  Meetings attract between 30 and 90 people, and are held at a restaurant or hotel in the bay area convenient for our speaker.  We usually convene at 6:00 pm for cocktails, dinner at 7:00 pm, and lecture at 8:15 pm.

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Last update: 9/3/2003