2014 Archived Content
Cambridge Healthtech Institute’s Sixth Annual
Engineering Genes, Vectors, Constructs and Clones
Upstream Decisions Lead to Downstream Success
January 13-14, 2014
Day 1 | Day 2 | Download Production Brochure | Speaker Biographies
TUESDAY, JANUARY 14
7:15 am Conference Registration
7:30 Breakfast Presentation: Leveraging Gene Synthesis for the Systematic Optimization of Protein Production
Mark Welch, Ph.D., Director, Gene Design, DNA2.0, Inc.
Current gene design and synthesis technologies allow control over sequence features critical for protein production. Variables at the levels of ORF coding, vector elements, protein sequence and the host genome can be essential for maximum system yield. We describe the application of gene synthesis and informatics technologies for the systematic study and optimization of such variables, with an emphasis on gene coding variables that influence protein yield in a number of commonly employed production hosts.
8:30 Chairperson’s Remarks
James E. Galen, Ph.D., Professor, Medicine, Chief, Salmonella Live Vector Vaccine Section, University of Maryland School of Medicine
8:35 Molecular Cloning, Overexpression and an Efficient One-Step Purification of αVβ5 and α5β1 Integrin
Lawrence J. Tartaglia, Ph.D., Research Scientist, Biochemistry and Molecular Biology, Center for Structural Biology, University of Florida
Recombinant αVβ5 and α5β1 integrin expression systems were created for the large-scale production of integrin extracellular domains that take advantage of Fos and Jun dimerization for expression in HEK293 cells and SF9 insect cells. Both integrin heterodimers were purified in a one-step nickel column purification scheme, characterized and yields were in quantities suitable for multiple applications including structural biology and functional assays.
9:05 Brevibacillus, a New Tool for High-Level Intracellular Expression of Bacterial Antigens
Domenico Maione, Ph.D., Unit Head, Cloning and Expression, Novartis Vaccines and Diagnostics
Brevibacillus choshinensis, a novel Gram positive Expression System, is an easy-to-handle non-sporulating bacterium. One major drawback that limits applicability is the absence of expression vectors based on inducible promoters. We achieved high level of intracellular protein expression in Brevibacillus, using the pHis1522 vector carrying the B. megaterium xylose-inducible promoter (PxylA). Our results suggest that the intracellular protein expression in Brevibacillus could be an attractive strategy to produce proteins that are poorly expressed, toxic or degraded in E. coli.
9:35 Selected Oral Poster Presentation: The Photosynthetic Bacterium Rhodobacter capsulatus as an Alternative Platform Organism for the Expression of Human Membrane Proteins
Armagan Özgür, Research Scientist, Institute of Molecular Enzyme Technology, Forschungszentrum Jülich, Heinrich-Heine University
The intricate nature of membrane proteins hampers their structural and functional studies because common expression hosts like E. coli are optimized for the production of soluble proteins. Therefore, we developed a new expression system based on the facultative phototrophic non-sulfur purple bacterium Rhodobacter capsulatus. Protein accumulation and localization studies revealed that E. coli seems to be the preferable expression host for human membrane proteins with low number of transmembrane helices, whereas membrane proteins with a higher number of transmembrane domains achieve higher protein yields with the newly developed R. capsulatus expression system. Therefore, the photosynthetic bacterium R. capsulatus is a promising alternative platform organism for the heterologous expression of more complex membrane proteins.
9:50 Coffee Break in the Exhibit Hall With Poster Viewing
10:50 A New and Versatile Concept for Systematic Multi-Gene Constructs Generation
Wassim Abdulrahman, Ph.D., Research Scientist, Mechanisms of Cancer, Friedrich Miescher Institute for Biomedical Research and CPC Novartis
Protein complexes expression is a major challenge for drug discovery. Although several molecular biology methods are well established for generating multi-gene constructs, none of these methods are adapted to in-parallel screening strategies. I will describe a novel and versatile method specifically designed for robotized screening of multi-protein expression.
11:20 Data Management and Automation of Protein Production Workflows in Biologics R&D
Christoph Freiberg, Ph.D., Senior Scientist, Biologics Research, Bayer HealthCare
Our comprehensive workflow platform supports highly parallelized antibody and tool protein production. It uniquely integrates in silico cloning, construct annotation and molecule registration functionalities, while supporting automated and high-throughput evaluation of expression systems of different scales. The molecule’s historical record is faithfully recorded, from early discovery campaigns to engineering and late-stage development. A single system stores molecule hierarchies and associated key expression, purification and analytics data. We thus glean essential information for improving product quality and optimizing production processes.
11:50 Sponsored Presentation (Opportunity Available)
12:20 pm Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own
4:45 Close of Conference
4:30-5:00 Short Course Registration
5:00-8:00 Dinner Short Courses (SC8-SC14) More Details >>
Day 1 | Day 2 | Download Engineering Brochure | Speaker Biographies