Co-Located with BIOTECHNICA 2009, Europe's No.1 Event in Biotechnology and Life Sciences 

Day 1 |  Day 2

WEDNESDAY, 7 OCTOBER

13:00 Conference Registration

 

Engineering a Better Expression Process

14:00-14:05 Chairperson’s Remarks

14:05-14:35 Development of a Novel Gateway-based Vector System for Efficient, Multiparallel Protein Expression in Escherichia coli, Insect and Mammalian Cells.

Felix Freuler, M.S., Scientific Technical Leader I, Center for Proteomic Chemistry, Novartis Institutes for BioMedical Research (NIBR)

We have generated a series of Gateway vectors which were improved with some new features: Kanamycin resistance for protein expression in E. coli, HRV 3C protease cleavage sites, novel solubilization tags and last but not least the capability to combine diverse N- and C-terminal extensions. This set of customized Gateway vectors proved to be highly useful for overexpression of a broad range of proteins in various hosts.

14:35-15:05 New Ways of Establishing Production Cell Lines for Structural Biology

Konrad Büssow, Ph.D., Project Group Leader, Structural Biology, Helmholtz Centre for Infection Research

Site directed, recombinase mediated cassette exchange (RMCE) has the potential of establishing stable mammalian producer cell lines fast and with reproducible expression strength and stability. We have established CHO master cell lines for RMCE for structural biology using a GFP vector and FACS sorting of high producer cells. A CHO Lec cell line was used, that is glycosylation deficient and produces glycoproteins that can be deglycosylated and crystallized efficiently. Establishing CHO production cell lines for target proteins by RMCE was fast (less than one month), reproducible and yielded cell lines that compare favorably to traditional production cell lines. Protein was produced by fermentation and crystals were obtained of deglycosylated, purified proteins.

15:05-15:35 Protein Science Technologies in Industrial Process Development

Philine Dobberthien, Process Science / Downstream Development, Boehringer Ingelheim Pharma GmbH & Co KG

This presentation will introduce novel technologies developed in the downstream development unit to accelerate process development and to enhance the knowledge and understanding of the behaviour of biomolecules we are handling. Downstream processes at Boehringer Ingelheim are designed to minimize the time to tox and time to clinical material, while maintaining a focus on the development of a customized process for each individual product. With the complexity of biological compounds and the increasing regulatory requirements, it is critical to have flexibility in the development process in order to insure the highest product quality and safety standards. Novel technologies based on biochemical and physical characterization, the implementation of automation and high-throughput techniques allow us to individually streamline process development timelines while increasing the amount of data used to make critical process decisions leading to rationally designed processes. This presentation will also describe how our novel technologies have impact on later stages of development and scale-up leading to integrated processes.

 

15:35-16:00 Refreshment Break

16:00-16:15 The ExoIN System: A Powerful Tool for Ectopic Protein Expression in Mammalian Cells
Konstantin Matentzoglu, CEO, Trenzyme Biotechnology
Clonal cell lines stably expressing a protein of interest are routinely used in the characterization of the potential physiological functions of mammalian proteins or for protein priduction, but the generation of such cell lines is laborious and resource consuming. Here, we made use of the fact that fusion proteins consisting of ubiquitin linked to the N terminus of a protein of interest are efficiently processed by ubiquitin-specific proteases into their respective free proteins within eukaryotic cells, and designed an expression system that allows easy and highly efficient selection of mammalian cells homogenously expressing a protein of interest by linking stoichiometrically the expression of a given target to an antibiotic resistance on one polyprotein. ExoIN enables us to rapidly generate stable homogenous populations of mammalian cells in as little as 2 weeks.

16:15-16:30 Sponsored Presentation (Opportunity Available)


16:30-17:00 Affinity Partitioning of Proteins Tagged with Choline-Binding Modules in Aqueous Two-Phase Systems

Isabel Velasco, Ph.D., Project Leader, Research and Development, Biomedal

We present a novel procedure for affinity partitioning of recombinant proteins fused to a choline-binding module (C-LytA or LYTAG) in an aqueous two-phase system (ATPS) containing poly(ethylene glycol) (PEG). Proteins tagged with the C-LytA module have affinity to the PEG-rich phase, whereas subsequent addition of the natural ligand choline specifically shifts their localization to the PEG-poor phase by displacement of the polymer from the binding sites. These systems show interesting advantages both for laboratory and industry as they are cost-effective, easy to scale up and suitable for continuous operation. Many variables can be manipulated to improve the partition, and compatibility with detergents allows the purification of membrane proteins. We have successfully purified diverse recombinant proteins with the choline binding polypeptide tag by affinity partitioning in ATPS. This process avoids some disadvantages of the solid affinity supports such us resin cost, preparation and recycling, column fouling or changes in the stability of the adsorbed proteins. The high purity degree from crude extracts in few simple steps (>98%), its cost-effectiveness and the easy scalability of the process make the affinity partitioning of proteins tagged with choline binding protein (LYTAG) a promising system for either basic research and industrial biotechnology.

17:00-17:30 Poster Spotlight Presentations

18:30 BIOTECHNICA Night: Beer Hall, full dinner reception, a traditional German band