PEGS Europe 2010 | Day 1 | Day 2
Great strides have been made in the expression of proteins for structure and characterization studies, as well as for therapeutics and diagnostics. However, with these strides have come new obstacles. Higher throughput expression and purification, as well as more flexible expression systems and techniques are in even greater demand now to support the needs of the research pipeline. It has become even more important to prevent problems in protein expression by wise choices in data analysis, vector choice/design, and cell line selection and expansion well before the task goes into the laboratory. This conference will explore the newest innovations relating to the expression of proteins for biological understanding and therapeutics, as well as the latest science in cell line development and optimization.
TUESDAY, 5 OCTOBER
9:00 Conference Registration and Morning Coffee
9:30 Chairperson’s Opening Remarks
9:35 KEYNOTE PRESENTATION
Protein Expression in Mammalian Cells: Past Perspective, Future Potential
John Birch, Ph.D., Consultant, Henley-on-Thames
More than half of all licensed recombinant therapeutic proteins are produced in mammalian cell systems. The development of efficient expression technologies, in combination with improved feeding strategies in fed-batch culture, has resulted in titres of grams per litre for well-expressed proteins such as antibodies. This talk will cover the key developments that led to this success and the challenges and opportunities that remain.
10:05 A Novel Fusion System for Soluble Overexpression of Recombinant Proteins in Escherichia coli
Sofia Costa, Researcher, Biological Engineering, IBBCEB, University of Minho
A gene fusion technology has been applied in the E. coli system. A novel and promising fusion system, consisting of two fusion tags – Fh8 and H tags, has recently been discovered and patented. Both fusion tags increased protein production yields in E. coli and may potentially promote a solubilization effect on difficult-to-express proteins with diagnostic/therapeutic application.
10:35 Coffee Break
11:00 Production of Recombinant Human Multi-Protein Transcription Factor Complexes
Arnaud Poterszman, Ph.D., Research Director (CNRS),Structural Biology and Genomics, IGBMC-CERMB
We present strategies for production of multi-subunit transcription factors such as nuclear hormone receptor complexes or the basal transcription/DNA repair factor TFIIH using the baculovirus expression system. Selected examples illustrate recent developments: (i) HTP mini expression screening, (ii) fluorescent proteins as markers and for quality control (iii) vector development for parallel cloning and (co-)expression of multiple constructs for a single target, (iv) single virus co-expression of multi-subunit complexes.
11:30 Minicircles (MCs): Overcoming the Limitations of Transient Expression Systems
Juergen Bode, Ph.D., Professor, Medical School Hannover
We discuss a ~4kb minicircle (MC), with superior nuclear transfer and expression characteristics due to its ccc-status. Following the initial expression phase MCs are able to exploit the cellular machinery to replicate in a way reminding of ARS-vectors. Several MCs can be established side-by side allowing the regulated expression of multi-subunit proteins. Genetic elements remaining in the minicircle (promoter, S/MAR, poly(A)-site) and the MC preparation procedure could continuously be refined.
12:00 How a Strong Expertise in Target Proteins Expression can Become an Added Value for Bioprocess Development
Hervé Ginisty, Ph.D., CSO, GTP Technology
In the field of recombinant protein expression, experience and know-how are often the key to success. During the past ten years, GTP has worked on more than 800 projects concerning the expression of over 400 challenging proteins.
In this presentation, we will outline how the large range of expression systems and purification strategies we have developed, allowed us to address most target proteins expression challenges, and now enable us to offer original and flexible solutions for bioprocess development projects.
12:15 From EnBase to EnPresso - Novel Instant Solutions Facilitate High Level Recombinant Protein and Plasmid Production
Peter Neubauer, Ph.D., Professor, Technische Universität Berlin, and Scientific Advisor, BioSilta Oy
Recently BioSilta have developed a new high cell density culture portfolio, EnBase®, which is based on a biocatalytically controlled release of glucose from glucose polymers. EnBase® can be applied in shaken cultures without any external feeding devices over a wide range of culture volumes and has proven its strength in many laboratories. New research focused on a further simplification of the practical usability of EnBase®, resulted in the development of the EnPresso™ product line, which contains tabletised or granulated sterile packed microbial cultivation media which can simply be added to pre-sterilized water. This product line paves the way for direct process scaling from microlitre to pilot-scale cultivations.
12:30 Lunch for Purchase in the Exhibit Hall
13:45 Dedicated Poster Viewing in the Exhibit Hall
14:30 Chairperson’s Remarks
14:35 A High-Throughput Microtiter Plate-Based Screening Method for the Detection of Full-Length Recombinant Proteins
Matthias Mack, Ph.D., Head, Institute for Technical Microbiology, Mannheim University of Applied Sciences
Escherichia coli is an important host for the production of proteins. The development/optimization of a protocol to overproduce a desired protein in E. coli is often tedious. A novel high-throughput screening method based on the Luminex® xMAP™ bead technology was developed allowing a rapid evaluation of a certain expression strategy. The new method was also applied to the analysis of proteins produced in Pichia pastoris and Pichia angusta.
15:05 Robo-Lector – A Novel Platform for Automated High-Throughput Cultivations in Microtiter Plates with High Information Content
Frank Kensy, Managing Director, m2p-labs GmbH
The presentation will introduce the novel RoboLector platform for automated microbial and cell culture cultivations. 48 or 96 parallel fermentations in a microplate format can be operated and manipulated by a liquid-handling robot triggered by non-invasive online monitoring signals such as biomass and fluorescent protein concentrations, pH and pO2. Data from a 2-D induction profiling of E.coli cultures expressing a FMN-binding fluorescent protein (FbFP) and several examples of media optimization will be presented.
15:35 Refreshment Break
16:00 Boosting Secreted Expression of Biologics from Pichia Pastoris with Novel Tools and Technologies
Thomas Purkarthofer, Ph.D., Head of Business Development, VTU Technology GmbH
Pichia pastoris represents an intriguing expression system for biologics production featuring notable expression and secetion power and cost and safety advantages. VTU applies its exclusive library of synthetic AOX1 promoter variants which trigger a substantial increase in productivity. A well characterized in-house expression toolset and specialized handling protocols ensure to shrink development timelines and ramp up speed-to-clinic. The company´s most recent development comprises novel AOX1 variants functioning without induction by hazardous methanol outperforming the (methanol-free) constitutive GAP promoter. Showcase examples applying this powerful Pichia system both in the presence and in the absence of methanol will be presented.
16:30 A Novel T7 RNA Polymerase-Dependent Expression System for High-Level Protein Production in the Phototrophic Bacterium Rhodobacter capsulatus
Thomas Drepper, Ph.D., Institute of Molecular Enzyme Technology (IMET), Heinrich-Heine-Universität Düsseldorf
17:00 Rapid Protein Quantification by Applying BioLayer Interferometry
Arnout Gerritsen, Director Assay & Bioanalytical Science, Genmab
17:30 Lactococccus lactis P170 Expression System: A Novel Secretion-Based Expression System for Production of Recombinant Proteins
Soeren Madsen, Ph.D., Group Leader, Bacterial Expression, Bioneer A/S
The P170 Expression system is based on the endotoxin-free Gram positive bacterium Lactococcus lactis. Gene expression is auto-induced during the transition to the stationary phase and the expression system is designed to secrete the recombinant proteins to the growth medium thereby making downstream processing more convenient. Examples will be given on production of recombinant proteins, which has entered phase II clinical trials.
18:15 Interactive Breakout Discussion Groups
19:15 – 21:00 CHI Networking Reception