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Baculovirus Technology - Day 1



Baculovirus Technology

Baculovirus vectors are widely used as tools for expressing proteins, delivering genes into cells, and creating vaccines. Once viewed as an alternative, the baculovirus system has achieved recognition through extensive clinical testing and regulatory exposure. The speed of reaching protein expression and the inability to transmit mammalian disease makes baculovirus an attractive platform.

The Baculovirus Technology meeting brings together experts who are innovating applications and achieving clinical success. Breakthrough approaches to tackling the problems of increasing protein expression levels including a session focused on BacMam will be featured, along with case studies from industry leaders.


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Wednesday, August 24

7:30 am Registration & Morning Coffee

 

UNDERSTANDING BIOLOGICAL PROPERTIES TO OPTIMIZE BACULOVIRAL MECHANISMS

8:25  Opening Chairperson’s Remarks

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

8:30  OPENING KEYNOTE PRESENTATION:
Baculovirus Nucleocapsid Transport to the Nucleus and Beyond!

Loy VolkmanLoy Volkman, Ph.D., Professor Emerita, Plant & Microbial Biology, University of California, Berkeley - Biography

Phylogenetic evidence indicates baculoviruses have been tracking their juvenile insect hosts for about 230 million years, since the radiation of the Lepidoptera. Lepidopteran larvae developed a formidable defense against pathogens during this period: an exoskeleton that covers their exterior surfaces including their respiratory tubes, foreguts and hindguts. Baculoviruses have countered this measure by targeting larval midgut columnar epithelial cells via their microvilli, an extraordinary biological feat.

9:00  FEATURED PRESENTATION:
Application of the Baculovirus Expression System in Generating Tools for Drug Discovery

Ian HuntIan Hunt, Ph.D., Associate Director, Discovery Technologies, Novartis Institutes for BioMedical Research, Inc.




9:30 Protein Glycosylation in the Baculovirus-Insect Cell System

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

The baculovirus-insect cell system is widely used to produce recombinant glycoproteins. However, the N-glycosylation patterns of these products typically differ from those of their native, mammalian cell-derived counterparts. For the past 15 years, my lab group has focused on insect cell glycoengineering to address this problem. This presentation will focus on our ongoing efforts to humanize protein N-glycosylation in the baculovirus-insect cell system.

10:00 Networking Coffee Break with Exhibit and Poster Viewing

 

VECTOR ENGINEERING

10:45 Baculoviral Vectors for Gene Editing of Human Stem Cells

Pin WangPin Wang, Ph.D., Associate Professor, Chemical Engineering, Biomedical Engineering, Pharmacy and Pharmaceutical Sciences, University of Southern California

We will describe our success of using a baculoviral vector (BV) system carrying zinc finger nucleases (ZFNs) for site-specific modification of cell genome. We observed that BV-mediated transient expression of ZFNs specifically disrupted the desired locus in transduced cells. When BV was engineered to deliver both ZFNs and donor DNA, permanent site-specific gene addition could be achieved.

11:15 Enabling Complex Protein Production in Academic and Industrial R&D

Imre BergerImre Berger, Ph.D., Group Leader, Structural Biology Unit, EMBL Grenoble - Biography

Complex proteins, involved in disease causing processes, are entering center-stage as key drug targets of the future. The baculovirus/insect cell expression system is particularly useful for producing such protein targets, biologics and multicomponent assemblies, for many applications. We have developed MultiBac, a BEVS designed for high-quality multiprotein complex production, and have installed MultiBac as a platform technology at the Eukaryotic Expression Facility (EEF) at the EMBL. Our MultiBac system and several of its successful applications, in academic and industrial R&D, will be presented.

 

Sponsored by
Blue Sky Bioservices 
11:45 Enhancements to Baculovirus Expression Technology

Norman Garceau, Ph.D., President & CSO, Blue Sky Biotech






12:00 pm Sponsored Presentation (Opportunity Available)

12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own

 

FOCUS ON BacMam

1:55 Chairperson’s Remarks

Daniel Fitzgerald, Ph.D., Eclosion SA

2:00 BacMam Gene Delivery in Drug Discovery

Robert AmesRobert S. Ames, Ph.D., Director, Cellular Targets, Biological Reagents and Assay Development, GlaxoSmithKline - Biography

BacMam gene delivery has gained prominence for the development of cell-based drug discovery assays due to the short cycle time for reagent generation but more importantly because of the unparalleled experimental flexibility offered by the technology. Rather than stable cell lines we instead generate recombinant BacMam viruses containing the cDNA of interest and use these virus preparations to transduce the target cells of interest. The presentation will highlight the versatility of the technology and how we have optimized and implemented it across drug discovery.

2:30 BacMam-Mediated Recombinant Protein Expression for Cell-Based Reagents

Kim EngwallKim Stutzman-Engwall, Ph.D., Associate Research Fellow, Pfizer Global R&D Groton Labs - Biography

Expression of recombinant protein in mammalian cells is a key step in the drug discovery process. Key advantages of recombinant expression in mammalian cells, include the appropriate signal transduction machinery and native glycosylation profiles.  These cells are routinely used for cell-based screening assays, binding assays, and selectivity assays. There are several options available for generating mammalian cells expressing recombinant proteins, including transient transfection, generation of stable cell lines and viral transduction using adenoviral, lentiviral, retroviral, or BacMam systems. While each option has advantages and disadvantages and a range of flexible options is desirable, we have recently focused our efforts on BacMam. Our investigations into the use of BacMam for mammalian cell transductions and the development of novel vectors have enabled us to efficiently optimize expression of poorly expressed proteins, as well as rapidly generate multiple isoforms of GPCRs for selectivity screening. Other key advantages of BacMam are the ability to ‘dial-in’ the desired level of recombinant protein expression as well as co-express multiple genes simultaneously. We have developed BacMam protocols to optimized large scale protein production using Wave Bioreactors. Several case studies using BacMam technology will be presented.

3:00 Baculoviruses for Lentivirus Generation

Kari AirenneKari Airenne, Ph.D., Professor, Biotechnology and Molecular Medicine, A.I. Virtanen Institute, University of Kuopio - Biography

We have studied possibility to produce lentiviruses by baculovirus technology. Our results show that safe 3rd generation lentiviruses can be produced by hybrid baculoviruses in adherent and suspension cells. High-titer LV stocks were readily gained in 293T cells and the viruses performed similarly as the LVs produced by conventional means in vitro and in vivo. A scalable and cost-effective capture purification step was also set up based on a DEAE monolithic column enabling of 65% recovery of highly purified lentiviruses.

3:30 Networking Refreshment Break with Exhibit and Poster Viewing

 

VACCINE DEVELOPMENT

4:15 Application of Baculovirus Technology in the Development of Sipuleucel-T, an Active Cellular Immunotherapy for the Treatment of Asymptomatic or Minimally Symptomatic Metastatic Hormone Refractory Prostate Cancer

Samuel LilSamuel Li, Ph.D., Associate Director, Pre-Clinical Discovery, Dendreon Corporation

Sipuleucel-T (PROVENGE®) is an FDA-approved autologous cellular immunotherapy for the treatment of asymptomatic or minimally symptomatic metastatic castrate resistant (hormone refractory) prostate cancer. This presentation will present the data on the clinical development of sipuleucel-T, and on our Antigen Delivery CassetteTM technology platform in general. Engineering and production of protein antigen PA2024 by the BEVS technology and sipuleucel-T manufacturing are also briefly described.

4:45 Production of Norovirus Virus-Like Particles in the Baculovirus Expression System from Benchtop to GMP Manufacturing

Ross TaylorRoss Taylor, Ph.D., Director, Process Development, LigoCyte Pharmaceuticals, Inc. - Biography

The baculovirus expression system has been used in both Wave and stirred-tank bioreactor systems for production of two distinct norovirus virus-like particles (VLP). Emphasis has been placed on employing single-use technologies including disposable bags for media storage and sample collection, disposable bioreactors for VLP production, capsule filters for VLP harvest, and membrane chromatography for VLP capture. LigoCyte’s VLP manufacturing process results in gram quantities of highly purified VLP which are currently being used in Phase I/II clinical trials.

5:15 Networking Reception, Last Chance for Poster and Exhibit Viewing

6:45 End of Day



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