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Module 3: Introduction to Proteomics and Protein Separation StrategiesFACULTY Scot R. Weinberger, President, CEO and Founder of GenNext Technologies™ Inc. Dr. Barry E. Boyes, Vice President of Biological Product Development, Smiths Detection Module three begins with a brief introduction to proteomics followed by specific discussion of protein purification strategies. The protein purification lecture will be organized around the dominant use of liquid chromatography (LC) separation methods for sample preparation, and for multidimensional separations prior to identification and characterization using mass spectrometry. Sample preparative methodologies covered will address common protein "wet chemical" approaches to dealing with sample biomass, including samples from various organisms and extracts of cells, tissues and biological fluids. Related topics of physical separations will be covered to understand contemporary proteomic sample workflow, and the options available for varying sample types, quantities, and experimental goals. Most critically, the sample compatibility considerations of different sample acquisition and processing methods for downstream LC and electrophoresis separations will be explored, as will be procedures for preserving sample integrity by reducing or controlling chemical and enzymatic alterations during processing. The various common LC methods for protein and peptide separations, and the physiochemical properties that underlie the use of these separation modes, will be presented as structure-retention models. Topics will include the most common operational modes: size exclusion, ion exchange, reversed-phase, hydrophobic interaction and affinity. Typical operational conditions covered will include mobile phase and stationary phase properties, linear velocity, temperature, and the impact of these choices on downstream processes. The application of these various modes will be demonstrated at the lab-preparative and analytical scales using modern HPLC instruments, columns, and software. The instrumental aspects of this section will include common LC methods, as well as the application of micro- and nano-scale formats for columns, instruments, plumbing, and MS interfacing. This presentation will review practical examples of coupling separations in both continuous and discontinuous transfer modes between separation dimensions. Examples described in detail will focus on protein identification and quantification in high complexity proteomic samples obtained from tissues and biological fluids. The final topics covered will address the use of affinity selection methods for preparative isolation and for proteomic sample complexity reduction. The focus in this discussion will be on the advantage of targeting a "chromatographically" defined sub-proteome, and to demonstrate the challenges faced by analysis of samples with extremes in target protein abundance.GenNext's 2007 In Person Translational Courses have been completed. GenNext would like to thank its Education Partners and excellent instructors for another highly received Translational Research Program.Information regarding GenNext's 2008 program will become available shortly. Sign-up to find out about upcoming GenNext courses. For more information about this course, please contact GenNext Technologies at inform@gennexttech.com or call 650-563-9577.
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