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10:00 am - 1:00 pm
(SC3) PROGRAM MANAGEMENT FOR SCIENTISTS
Congratulations! You have been promoted to Program Manager. Now what? Can your bench-top scientific experience in proteins - expression, characterization, development of protein-based therapeutics, bioprocessing, etc. - lead to success in people/business management? Most scientists train for years in their prospective fields but must learn business management “on-the-fly.” This course is designed for scientists who have recently been promoted to program/product manager or are planning ahead.
Course Instructor:
 Patricia Seymour, M.B.A., Senior Consultant, BioProcess Technology Consultants
Patricia Seymour has over 20 years of experience in the biotechnology industry. Her expertise spans the development and commercialization spectrum, including biologics and small molecule development and manufacturing, supply chain strategy and management and operations leadership. Her current activities include: developing overall CMC strategies for diverse product pipelines, developing and implementing outsourcing strategies, managing outsourced process development and manufacturing activities from drug substances through to distribution, steering clients through CMC regulatory requirements and overall CMC operations including project management and budgeting. Prior to joining BioProcess Technology Consultants, Ms. Seymour was Senior Director, Global Investigational Supply Operations at Millennium Pharmaceuticals, Inc., where she worked with other senior leaders to develop and implement CMC strategies including outsourcing. She was previously Director of Business Development at Covance Biotechnology Services (now Diosynth) where her responsibilities included negotiating contact manufacturing projects. As Senior Director, Corporate Development at Collaborative BioAlliance she was instrumental in launching the new contract manufacturing business unit. Before Collaborative, Ms. Seymour held research and development positions at ImmunoGen, Dana Farber and Sloan Kettering. Ms. Seymour received her B.S. from Villanova University and her M.B.A. from Boston University. Ms. Seymour is Certified Supply Chain Professional (APICS).
2:00 pm - 5:00 pm
(SC8) MEMBRANE PROTEINS - An Important Protein Class
This course will provide an overview of Membrane Proteins; their structure, function, interactions, and the inherent challenges of working with their hydrophobic natures. Like a new frontier, Membrane Protein structures and functions are continually being discovered and revealed. They offer insights into further understanding protein interactions, and tantalizing promise as disease targets. The Membrane Proteins Short Course is the perfect overview for those familiar with soluble proteins who want to expand their protein repertoire.
2:00 Membrane Proteins: Life in an Oily Phase
William A. Cramer, Ph.D., Henry Koffler Professor, Biological Science, Purdue University
This talk will provide an overview of Membrane Proteins:
1. State of the field
2. Problems: hydrophobicity mismatch; expression; extraction; purification (stability; proteolysis; delipidation; activity); hetero-oligomericity
3. Crystallization: 3D (in surfo, in meso, in bicelle) for analysis by x-ray diffraction; 2D, electron diffraction.
4. Crystal analysis: recent technical development; synchrotron beam lines measure small (< 10 μ) crystals.
5. Survey of membrane protein systems: GPCR, transporters, energy transducers, receptors, α-helical vs. β-barrel outer membrane proteins.
2:45 Establishing a Structure Based Drug Design Loop for the CGRP Receptor, a Class B GPCR
Ernst ter Haar, Ph.D., Research Fellow I, Structural Biology, Vertex Pharmaceuticals, Inc.
The calcitonin gene-related peptide (CGRP) is a potent vasodilator directly implicated in the pathogenesis of migraine. Its receptor (CGRP-R) is a heterodimer containing the calcitonin receptor-like receptor (CLR), a class B GPCR, and RAMP1, a receptor activity-modifying protein. In order to aid our drug design effort against CGRP, we solved the crystal structure of the CGRP-R heterodimer ectodomain and used it for structure-based design of novel antagonists. We also obtained the structures of CLR/Ramp1 in complex with olcegepant (BIBN4096BS) and telcagepant (MK0974), two clinically validated antagonists that provided the proof of concept for CGRP as a migraine target. The inhibitor complex structures from our effort helped us develop a pharmacophore model for the CGRP-R ectodomain and explore starting points for inhibitor design.
3:30 Refreshment Break
4:00 Transporters: the Workhorse Machines of Biological Membranes
Robert K. Nakamoto, Ph.D., Professor and Vice Chair, Department of Molecular Physiology & Biological Physics, University of Virginia
Transporters carry out the work of importing nutrients, extruding toxic compounds, and forming ionic gradients. They are involved in energetic processes such as neuronal transmission, photosynthesis and oxidative phosphorylation. When the transport mechanism is coupled to energy, they are among the most efficient machines in Nature. They have long been the subjects of intense studies because of their importance in cellular physiology, as key components of basic cellular functions, and as potential drug targets. The biochemistry and structural biology of transport proteins is almost always difficult because of they are generally large integral membrane proteins that usually work as oligomers or have multiple subunits. Because of the transport function, the proteins have multiple conformational states and are often flexible and highly dynamic. Understanding of the biochemical and biophysical properties of these proteins has been instrumental in obtaining crystals and eventually solving their high-resolution structures. In some cases, the molecular details of the conformational states during the transport cycles show that the proteins undergo large-scale domain movements during the transport cycles. The transporters are true molecular machines that carry out the work of moving solutes across membranes.
- Roles of transporters in cellular functions
- Transporters define cellular physiology and bioenergetics
- Contrasts between transporters, channels and porins
- Types of the transporters: pumps, secondary transporters, facilitated diffusion
- The biophysics and biochemistry of transporters
- The structural biology of transporters
- How transporters work
- Transporters in human disease
- Transporters as drug targets and their roles in multiple drug resistance
5:00 Close of Membrane Proteins Short Course
5:30 pm – 8:30 pm
(SC10) AFFINITY TAGS FOR PROTEIN PURIFICATION
Dinner, Presentations & Interactive Discussion
- Overview of Affinity Purification
- Types of Affinity Tags
- Comparison of Affinity Tags
- Tag Cleavage/Self-Cleaving Tags
- Alternative Tagging Strategies
- Streamlining Protein Recovery
- Emerging Technologies
5:30 Chairperson's Remarks
John Proctor, Ph.D., Product Manager, Biosensors and Assay Kits, FortéBio, Inc.
5:40 Choosing the Right Tag: For You and Your Protein
 William Gillette, Ph.D., Senior Scientist, Protein Expression Lab, SAIC-NCI Frederick
Affinity tags offer a wide array of features and drawbacks for both protein expression and purification. A discussion of the pros and cons of these tags will be followed by examples of how their use simplifies (or complicates) protein expression and purification in a Core Service lab.
6:45 Q&A -- Discussions
7:00 Dinner Break
7:30 Using Tandem Affinity Purification-Mass Spectrometry for Analyzing Intracellular Signaling Complexes
 Alexey Veraksa, Ph.D., Assistant Professor, Biology Department, University of Massachusetts, Boston
In this talk, I will use a case study of mapping the protein complexes in the Notch pathway to share our experiences with tandem affinity purification (TAP)-mass spectrometry approaches. The Notch signaling pathway is a conserved mechanism of intercellular communication that is used in all metazoans to establish molecular differences between adjacent cells. We have used the original TAP tag, as well as more recently developed GS-TAP tag, to analyze protein interactions in the Notch pathway. We have validated some of the novel interactions using functional assays in cultured cells and in vivo. I will discuss the advantages and pitfalls associated with the use of the TAP tags, and will share our new data on developing even more efficient ways to isolate and analyze protein complexes.
8:30 Close of Affinity Tags for Protein Purification Short Course
Speaker Bio Sketches
Dr. William Gillette
The Protein Expression Laboratory provides protein expression/purification core service to the laboratories of the NCI, NIH and USAMRIID. Dr. Gillette is currently focusing on micro-scale purification and supporting analysis techniques to evaluate protein expression constructs and the success of the micro-scale chromatography experiments. Dr. Gillette earned his Doctorate in Microbiology from North Carolina State University, Raleigh, North Carolina.
Dr. Alexey Veraksa
My laboratory studies signaling networks in Drosophila development, with a focus on dynamic protein-protein interactions that control developmental signaling pathways. As a postdoctoral fellow at the MGH Cancer Center, Harvard Medical School, I applied new protein purification methods in Drosophila and used them in combination with mass spectrometry to analyze protein complexes in the Notch interaction network. This study identified the Drosophila β-arrestin Kurtz (Krz) as a Notch signaling modulator that regulates the turnover of the Notch receptor. After becoming an independent researcher at UMass Boston, I continue to investigate the diverse functions of β-arrestin Krz in Drosophila development. We have recently found that Krz inhibits MAPK signaling at multiple developmental stages by a previously unknown mechanism. Part of our research efforts is devoted to developing new protein purification strategies that can be used for isolating and characterizing protein complexes involved in signal transduction. I received my Doctorate in Developmental Biology from the University of California, San Diego in La Jolla, California.
*Separate Registration Required
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