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Many proteins pose extreme challenges in expression due to their particular properties. Membrane proteins, toxic proteins, proteins which don’t fold correctly when expressed, or express in insoluble forms continue to cause hardships for protein researchers. The 5th Annual “Solving Difficult Protein Problems from Expression through Purification” will provide cutting-edge solutions to these challenges with the latest strategies from researchers who have solved these problems.
WEDNESDAY, 12 OCTOBER
13:00 Conference Registration
14:00 Chairperson’s Remarks
Darren J. Hart, Ph.D., Grenoble Outstation, European Molecular Biology Laboratory
14:05 Engineering Mammalian Cell Lines and Vectors for Improved Secretion of Hard to Express Proteins
Nic Mermod, Ph.D., Director, Institute of Biotechnology, University of Lausanne
Methods to increase transgene integration using a gene transfer process based on homologous recombination will be presented, opening new avenues to engineer cells to achieve more efficient recombinant protein expression. Protein secretion is another bottleneck, especially for difficult to express secreted proteins. We will illustrate how the faulty step can be identified and show that overexpression of specific combinations of secretion pathway proteins may solve processing and secretion issues for difficult to express proteins.
14:35 Random Library Approaches for Expression of Challenging Proteins
Darren J. Hart, Ph.D., Grenoble Outstation, European Molecular Biology Laboratory
ESPRIT is a directed evolution-type method that permits solubility screening of tens of thousands of random constructs of a single target gene. Expression and purification of domains from many challenging, poorly annotated proteins has been achieved for applications in structural biology and immunisation. The core technology plus recent developments for studying protein complexes and increasing throughput will be presented.
15:05 Humanization of the Plant N-glycosylation Pathway for the Production of Therapeutically Relevant Proteins
Herta Steinkellner, Ph.D., Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences
The presentation focuses on the manipulation of the plant N-glycosylation pathway for the production of recombinant proteins with a defined N-glycosylation pattern. We demonstrate knock in and knock out approaches to generate recombinant proteins with a customized N-glycosylation profile, including sialylation, the most complex human type of glycosylation. The impact of N-glycosylation to the function of recombinant proteins will be discussed.
15:35 Refreshment Break - Networking with Sponsors
16:15 Sponsored Presentations (Opportunities Available)
16:45 The MultiBac Platform at EMBL: Enabling Complex Protein Expression in Academic and Industrial R&D
Imre Berger, Ph.D., The European Molecular Biology Laboratory EMB
The baculovirus/insect cell expression system is particularly useful for producing such protein targets, biologics and multicomponent assemblies, for many applications. We have developed MultiBac, a BEVS designed for high-quality multiprotein complex production, and have installed MultiBac as a platform technology at the Eukaryotic Expression Facility (EEF) at the EMBL. Our MultiBac system and several of its successful applications, in academic and industrial R&D, will be presented.
17:15 High Yields of Functional Soluble T Cell Receptors after Stability Engineering
Geir Åge Løset, Ph.D., Research Associate, Department of Molecular Biosciences, Centre for Immune Regulation, University of Oslo
The vast repertoire of T cell receptors manifests the host diversity orchestrating adaptive immunity. Their detailed characterization is often challenging, as they express poorly as soluble molecules. We have systematically improved intrinsic molecular stability by homology defined point mutations, domain format variants and phage display engineering resulting in high yields of functional soluble T cell receptors.
PLENARY KEYNOTE SESSION
18:00 Keynote Introduction
18:05 Protein Engineering: Benefiting Therapeutic Proteins and Small Molecule Drugs Alike
 Andreas Plueckthun, Ph.D., Professor, Biochemical Institute, University of Zurich
18:40 'Systems Patientomics': The Future of Medicine
Hans Lehrach, Ph.D., Director & Head, Vertebrate Genomics, Max Planck Institute for Molecular Genetics
Ten years after the completion of the human genome in a ten year international collaboration at a cost of between 1 and 3 billion Dollar, we are now getting ready to be able to sequence genomes/ transcriptomes as part of routine medical practice in oncology. The flagship project IT Future of Medicine would extend this approach to generate integrated anatomical/molecular models of every patient in the healthcare system, as the basis for a data rich, computation intensive, individualized medicine of the future.
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19:15 – 21:00 CHI Networking Dinner Reception