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Baculovirus Technology - Day 2



Baculovirus Technology 


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Thursday, August 25

7:30 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee

 

PROTEIN EXPRESSION 

8:25 Chairperson's Remarks

Don JarvisDonald Jarvis, Ph.D., Professor, Molecular Biology, University of Wyoming






 

8:30 Featured Presentation:

Virion-Free Protein Production from Baculovirus-Infected Insect Cells

Just VlakJust M. Vlak, Ph.D., Professor, Vice-Dean and Member Academic Board, Laboratory of Virology, Wageningen University - Biography 

A novel genetic strategy has been developed to manufacture biopharmaceuticals in insect cells with baculovirus vectors, but without the presence of contaminating baculovirus particles (virions). Rigorous purification procedures are often needed to remove or eliminate particles from the biopharmaceutical products. This novel strategy greatly simplifies the downstream processing of biopharmaceuticals produced in insect cells, hence reduces production costs and enhances the safety of baculovirus-insect cell produced biopharmaceuticals for vaccines and gene therapy.
 

9:00 Tools for Improving Protein Production in the Baculovirus System

Dom EspositoDominic Esposito, Ph.D., Principal Scientist, Protein Expression Laboratory, SAIC-Frederick, Inc.

Our lab has developed a robust microscale protein production platform which allows us to rapidly analyze a large number of variables to optimize protein expression. Utilizing this system for baculovirus expression, we have been able to improve our chances of successful protein production using a combination of promoters, fusion tags, strains, and expression conditions. The platform technology and its application to the BEVS will be discussed and results of a number of experimental tests of the system will be presented.
 

9:30 Optimizing Efficiency and Effectiveness when Producing Proteins with the Baculovirus Expression Vector System

Krista Bowman, Ph.D., Senior Scientific Manager, Structural Biology, Genentech, Inc.

We have implemented a fairly high-throughput generic approach to increase not only our efficiency but also our effectiveness in successfully producing proteins with the baculovirus expression vector system. We have combined techniques such as small scale baculovirus infections in multiwell blocks, analysis of expression level following single affinity capture and elution from Ni-NTA Phytips, and analytical fluorescent size exclusion chromatography to gain insight into protein aggregation, multimerization, or complexation. Combining these semi-quantitative and qualitative approaches, we can more rapidly prioritize constructs for further characterization and production.

10:00 Networking Coffee Break

 

PROTEIN PRODUCTION 

10:30 Production of Recombinant Human Multi-Protein Transcription Factor Complexes

Arnaud Poterszman, Ph.D., Research Director, Integrated Structural Biology, IGBMC CNRS/Inserm/Universite de Strasbourg

We present here our strategies for the production of multi-subunit human transcription factors using the baculovirus expression system. Selected examples illustrate recent developments and their impact on the structural biology of human protein: (i) HTP mini expression screening, (ii) use of fluorescent proteins as makers 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.

11:00 Production of Mammalian Vectors in a Non-Mammalian System

Arie OorschotArie van Oorschot, Scientist, Upstream Processing, Process Development, Amsterdam Molecular Therapeutics (AMT) - Biography 

Potential commercialization of rAAVs produced in mammalian cells is hampered by the inability to produce rAAVs at large scale at acceptable costs. An attractive alternative is the baculovirus expression vector system, a non-mammalian production system by means of baculovirus expression vectors and insect cell suspension cultures. In this scalable production system, the essential components are delivered and assembled into rAAVs using infection of insect cells by three types of baculoviruses.
 

11:30 Addressing the Scale-Up Problem of Baculovirus Expression

Daniel FitzgeraldDaniel Fitzgerald, Ph.D., Eclosion SA - Biography 

We have addressed the "scale-up problem" by developing a novel baculovirus expression system for insect cells, called "iBac", in which recombinant protein expression is turned "off" during scale-up, and is turned "on" only when sufficient recombinant viruses are available for the desired expression scale, resulting in significant improvements in performance during scale-up relative to previously available systems.

 

12:00 pm End of The Bioprocessing Summit



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