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The availability of crystal structures is allowing greater understanding of membrane proteins, including ion channels and GPCRs. Phage and yeast display are being adapted for use with membrane proteins. Techniques for selection, co-crystallization, agonist binding and activation, expression and purification of proteins will be investigated. These will be exploited for drug development against difficult targets.
RECOMMENDED SHORT COURSES*:
Digital PCR Applications and Advances
*Separate registration required.
7:30 am Registration and Morning Coffee
8:25 Chairperson's Opening Remarks
8:30 Phage and Yeast Display Libraries and Screening
Andrew M. Bradbury, M.B. B.S., Ph.D., Staff Scientist, Biosciences, Los Alamos National Laboratory
James D. Marks, M.D., Ph.D., Professor, Anesthesia & Pharmaceutical Chemistry, University of California, San Francisco; Chief of Anesthesia and Vice Chairman, Anesthesia & Perioperative Care, San Francisco General Hospital
The session will bring scientists up to speed on the latest tools available for display technologies and is tailored for both the novice and those with experience in display technologies. The session will provide an overview not exclusive to membrane proteins, including phage display and construction of phage-displayed peptide, scFv and Fab libraries, yeast display and construction of yeast-displayed scFv and Fab libraries, selection and screening technologies that are compatible with phage and yeast-display libraries.
10:00 Networking Coffee Break with Poster Viewing
10:30 Phage and Yeast Display Libraries and Screening (Continued)
Andrew M. Bradbury, M.B. B.S., Ph.D., Staff Scientist, Biosciences, Los Alamos National Laboratory
James D. Marks, M.D., Ph.D., Professor, Anesthesia & Pharmaceutical Chemistry, University of California, San Francisco; Chief of Anesthesia and Vice Chairman, Anesthesia & Perioperative Care, San Francisco General Hospital
12:00 pm Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own
1:25 Chairperson's Remarks
1:30 Expressing Membrane Proteins in Yeast and Other Eukaryotic Cells
Robert M. Stroud, Ph.D., Professor, Biochemistry & Biophysics, University of California, San Francisco
This presentation addresses the key issues that need to be addressed to obtain pure, functional, homogeneous, stable membrane proteins as well as lessons and methods that are advancing this key area of membrane biology in the service of human health.
2:00 Selection of Internalizing Phage Antibodies Using Tumor Cells and Yeast Displayed Tumor Antigens
James D. Marks, M.D., Ph.D., Professor, Anesthesia & Pharmaceutical Chemistry, University of California, San Francisco; Chief of Anesthesia and Vice Chairman, Anesthesia & Perioperative Care, San Francisco General Hospital
We show that such antibodies to specific tumor antigens can be generated by first selecting phage antibody libraries on a tumor cell line expressing the target antigen followed by selection on yeast displaying the same antigen on their surface. Advantages and specific examples will be covered.
2:30 Filamentous Phage Display Vectors for Different Protein Classes
Andrew M. Bradbury, M.B. B.S., Ph.D., Staff Scientist, Biosciences, Los Alamos National Laboratory
3:00 Networking Refreshment Break with Poster Viewing
3:30 Adapting Yeast Antibody Display for Membrane Protein Targets
Eric V. Shusta, Ph.D., Associate Professor, Chemical & Biological Engineering, University of Wisconsin, Madison
Membrane proteins are challenging to work with in terms of antibody selection, engineering, and antigen identification as a result of their insolubility in aqueous solutions. We have therefore developed a platform for antibody engineering using either whole cells or cell lysates as antigen sources.
4:00 Antibodies against GPCR Dimers as Tools for Exploring Tissue Distribution, Regulation and Mapping Functional Domains of Receptor Heteromers
Lakshmi A. Devi, Ph.D., Professor, Pharmacology & Biological Chemistry, Mount Sinai School of Medicine
Mounting evidence suggests that GPCRs form dimers and dimerization is necessary for receptor maturation, signaling and trafficking. Using GPCR heterodimer-selective antibodies we have begun to characterize tissue localization, regulation and heteromer-specific function as well as to map their functional domains.
4:30 Cell-Free Expression of Polytopic Membrane Proteins for Drug Research
Frank Bernhard, Ph.D., Lab Leader, Institute of Biophysical Chemistry, Goethe University Frankfurt
This presentation demonstrates the systematic development of expression protocols for the cell-free production of high quality membrane proteins in preparative scales. The quality of the cell-free expressed proteins is demonstrated by a number of functional and structural approaches.
5:00 Close of Day