Day 1 | Day 2
The discovery and expression of proteins for structural studies, therapeutics and diagnostics continues to be a growing field of endeavor. The increased demands for larger quantities of protein in a shorter time leads to both problems and solutions. CHI’s 5th Annual, “Enhancing Expression and Achieving Higher Throughput through Cell Line Development” presents the newest strategies for overcoming these challenges, along with Case Studies from successful researchers who have solved these problems in creative and practical ways.
TUESDAY, 11 OCTOBER
9:00 Conference Registration and Morning Coffee
9:30 Chairperson’s Opening Remarks
Trevor Wilkinson, Ph.D., Associate Director, Antibody Discovery and Protein Engineering, MedImmune
9:35 Transient and Stable Mammalian Expression Platforms to Support Antibody Drug Discovery
Trevor Wilkinson, Ph.D., Associate Director, Antibody Discovery and Protein Engineering, MedImmune
We have developed a number of expression platforms using a combination of transient and stable cell line expression systems. These are used to produce protein antigens to facilitate the discovery of potential antibody therapeutics. The presentation will describe these systems and provide some case studies of their application.
10:05 Optimizing Protein Pipeline for Drug Discovery
Janet Sim, Ph.D., Investigator III, Oncology Protein Science, Novartis
To support drug discovery from target validation, assay development and crystallography the Oncology Protein Science group has implemented a flexible process and built optimized expression tools. This includes a suite of technologies to enable rapid testing of host systems (mammalian, insect and E. coli), promoter screens, vector optimization, rapid protein detection tools, chromatography optimization and knowledge-based protein truncation design. I will discuss our platform and use various examples to discuss strategies to overcome challenges for generation of protein and protein complexes.
10:35 Coffee Break - Networking with Sponsors
11:15 Sponsored Presentation (Opportunity Available)
11:45 Artificial Induction of the Unfolded Protein Response: A Tool to Increase Protein Production in Pichia pastoris
Mouna Guerfal, M.Sc., Researcher, Unit for Medical Biotechnology, Laboratory for Protein Biochemistry and Biomolecular Engineering, VIB Department for Molecular Biomedical Research, UGent
Artificial induction of the unfolded protein response (UPR) by overexpression of the transcription factor Hac1p can be used to improve the yield of recombinant proteins. After characterization of the transcription factor, Hac1p overexpression was evaluated on the production of secreted, surface displayed and membrane proteins. We show that the homogeneity and the expression levels of heterologous proteins can be increased by co-expression with Hac1p.
12:15 Sponsored Presentations (Opportunities Available)
12:45 Lunch for Purchase in Exhibit Hall 9
13:45 Dedicated Poster Viewing in Exhibit Hall 9
14:30 Chairperson’s Remarks
Michael R. Dyson, Ph.D., Senior Research Associate, Biochemistry, University of Cambridge
14:35 Transient Production of Recombinant Antibodies and Fusion Proteins in HEK293-6E Cells
Thomas Schirrmann, Ph.D., Research Group Leader and Lecturer, Department of Biotechnology, TU Braunschweig
Optimized antibody phage display can accomplish the demands of upcoming affinity proteome projects. We use the IgG-like scFv-Fc antibody format to address many application issues. To overcome the production bottleneck we improved a transient protein expression system using human embryonic kidney HEK293-6E cells allowing volumetric yields of more than 400 mg/L for a large number of different antibodies.
15:05 Selection of Soluble Protein Expression Constructs: The Experimental Determination of Protein Domain Boundaries
Michael R. Dyson, Ph.D., Senior Research Associate, Department of Biochemistry, University of Cambridge
Where expression of a full-length protein is problematic, it is often useful to clone and express individual protein domains. However, the annotated domain boundaries in databases such as Pfam or SMART are not always accurate. Various rational and combinatorial strategies for the experimental determination of the true protein domain boundaries will be described, including biophysical methods to screen selected clones.
15:35 Refreshment Break - Networking with Sponsors
16:15 Sponsored Presentation (Opportunity Available)
16:45 Stepwise Engineering of a Pichia pastoris D-amino Acid Oxidase Whole Cell Catalyst
Anton Glieder, Ph.D., Professor of Biotechnology, Austrian Centre of Industrial Biotechnology, Graz University of Technology
Based on a new open source Pichia pastoris expression platform advanced expression technologies were developed employing additive and synergistic advantages from gene design, protein targeting and strain engineering. Examples including soluble and membrane bound intracellular enzymes as well as secreted biocatalysts will be shown as examples where these new tools and technologies have been applied.
17:15 Pichia pastoris is Superior to E. coli for the Production of Recombinant Allergenic Non-Specific Lipid-Transfer Proteins
Martin Himly, Ph.D., Christian Doppler Laboratory for Allergy Diagnosis and Therapy, University of Salzburg
The non-specific lipid-transfer protein (nsLTP) of hazelnut represents a clinically important allergen termed Cor a 8. For production of a properly folded and biologically active recombinant Cor a 8 several expression systems were tested and the recombinant molecules characterized in detail. P. pastoris seemed superior to E. coli to obtain large quantities of soluble, properly folded, and biologically active rCor a 8.
17:45 The Challenge of Making Oxidation-Sensitive Proteins: Recombinant Production of an RNase Inhibitor
Peter Neubauer, Ph.D., Laboratory of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin
Here we describe different approaches for production of active RI in Escherichia coli, which include (a) production as a fusion protein [1], production as an authentic RI in the periplasm, as well as production of authentic RI in the E. coli cytoplasm. For all strategies fed-batch processes were developed which result in a high production of the target protein. The actual study shows, how physiological knowledge combined with high throughput screening strategies can be successfully applied for a straight forward bioprocess development also for difficult-to-produce considered proteins.
18:15 Interactive Breakout Discussion Groups
19:15 BIOTECHNICA EVENT NIGHT - Keynote Presentations followed by Networking Reception, Live Music and Dancing