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Arrive early and attend The Fourth Annual
Optimizing Cell Culture Development
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September 15-16, 2008
Boston, MA


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Baculovirus Technology - Day 2


Conference Proceeding CD Now Available
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THURSDAY, SEPTEMBER 18 - DAY TWO

8:00am Morning Coffee

DRUG DISCOVERY & DESIGN

8:25 Chairperson’s Remarks

8:30 Baculovirus Protein Expression Strategies for Drug Discovery
Kerry Kelleher, MSCIS, Manager, Baculovirus Expression, Senior Research Scientist II, Wyeth Research
The oral presentation will emphasize a rational approach to Baculovirus expression to achieve high throughput screening (HTS) and structural biology objectives. A protein production process paradigm including construct design, expression optimization, and then scale-up to increase protein production levels will be presented. In addition, a case study will highlight an information-guided strategy to Baculovirus protein expression to support both HTS and crystallography efforts.

9:00 RNA Interference to Enhance Cell Performance
William Bentley, Ph.D., Robert E. Fischell Distinguished Professor & Chair, Fischell Department of Bioengineering, UMCP, Center for Biosystems Research, UMBI, University of Maryland, College Park

9:30 Parallel Protein Production in the Baculovirus Expression Vector System for Structure-Based Drug Design
Ciarán N. Cronin, Ph.D., Head, Parallel Protein Production Group, Pfizer Global R&D
Pfizer’s La Jolla Laboratories have responsibility for the company’s small molecule drug discovery efforts in oncology, using the approach of structure-based drug design. Many oncology targets for drug intervention are protein kinases that are involved in signal transduction pathways. Production of this class of proteins in recombinant form has traditionally relied heavily on the baculovirus expression vector system (BEVS). The Parallel Protein Production Group at Pfizer’s La Jolla campus utilizes a highly parallel and generic approach to the cloning, expression and purification of recombinant protein kinases, and other oncology protein drug targets, that are suitable for co-crystallization prior to structure determination. This presentation describes the features of this parallel approach with particular emphasis on the involvement of the BEVS system for recombinant protein production.

10:00 Multi-Parallel Protein Production: Overcoming the Challenges to Accelerate Structural Based Drug Design
Ian Hunt, Ph.D., Associate Director, Discovery Technologies, Protein Structure Unit,
Center for Proteomic Chemistry, Novartis Institutes for BioMedical Research Inc.
For most biotech and pharmaceutical companies’ protein production groups play a crucial role in the drug discovery process. Specifically, the recombinant proteins they generate contribute to several key stages of the drug discovery pathway and which include exploratory research, target validation, high throughput screening (HTS) and structural biology studies. Therefore the quick and rapid production of high quality recombinant protein is a crucial component of many a drug discovery campaign. In an effort to enhance the speed and throughput of the system and thus align it to the demands of the modern drug discovery environment, significant efforts have been made by many groups to streamline the process to enable faster delivery of recombinant protein. This presentation will therefore use a series of case studies from a number of different drug targets to illustrate the utility of a multi-parallel baculovirus mediated protein expression (in the context of supplying proteins for structure based drug design) in achieving this goal. The presentation will also include discussion on some of the current bottlenecks within the process and potential new and enabling technologies that may assuage them.

10:30 Networking Coffee Break

PROTEIN PRODUCTION

11:00 Development of a Bench Scale-Up/Scale-Down Model for Virus Like Particle (VLP) Vaccine Production in WAVE and Other Disposable Bioreactors
Rahul Chelikani, Ph.D., Scientist, Process Development, Novavax
This talk will address crucial aspects about scale up and manufacturing of vaccines using a baculovirus-insect cell system. Discussion on key process control parameters like oxygen uptake rates (OUR), pH, temperature and metabolite consumption rates to be considered for optimal VLP production. Practical design and characterization of a 2L bench scale bioreactor model for process optimization and scale-up to WAVE bioreactors and disposable stirred tank bioreactors.

11:30 Improved Recombinant Protein Production by ParaTechs’ Vankyrin-Enhanced Baculovirus Expression Technology
Jeremy Kroemer, Ph.D., Senior Research Scientist, ParaTechs Corp.
ParaTechs Corp. has identified a gene family (vankyrins) from an insect virus that significantly delays death and lysis of baculovirus infected cells while enhancing recombinant protein production. ParaTechs’ vankyrin-enhanced BEVS (VE-BEVS) increased recombinant protein production up to 15 fold when an intracellular protein or a secreted protein were coexpressed with the vankyrin protein from a dual BEVS (VE-BEVS). When monoclonal Sf9 insect cell lines stably expressing vankyrin protein were used to provide the protein activity in trans, a 5-fold increase in intracellular protein production and up to 9-fold increase in secreted protein production was obtained. As with VE-BEVS, an increase in cell viability and prolonged protein expression post-infection was also observed with VE cell lines. Furthermore, VE transfer vectors, VE baculovirus DNA and additional VE insect cell lines are in the late stages of development.

12:00pm Development and Scale-Up of Protein Production Using the Baculovirus Expression System
Divya Parekh, Senior Process Engineer, Process Sciences, Diosynth Biotechnology, a part of Schering-Plough Corporation
The talk will outline the development and scale-up of a process for manufacturing protein in the BEVS. Implementation of the BEVS in a manufacturing environment requires balancing various scientific, regulatory, and economic issues. Key considerations and solutions for the scale-up of the process from the bench to large-scale bioreactors using the BEVS system will be discussed.

12:30 End of Baculovirus Technology 

SCIENTIFIC ADVISORS
Manon MJ Cox, Ph.D., Chief Operating Officer, Protein Sciences Corp.
William R. Hermans, Manager, Cell Culture, Blue Sky Biotech Inc.
Donald L. Jarvis, Ph.D., Professor, Molecular Biology, University of Wyoming

 



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