Sunday, September 24

4:00 - 6:00 Early Conference Registration

Monday, September 25 - Day One

7:30 - 8:30 Registration and Morning Coffee

Baculovirus: The Big Picture

8:30 - 8:40 Chairperson's Remarks 

9:20 - 10:00 FEATURED PRESENTATION

Insect Cell Cultures as Protein Factories: Status and Challenges Robert R. Granados, Ph.D., Virologist and Charles E. Palm Scientist, Emeritus, Boyce Thompson Institute for Plant Research, Cornell University Dr. Granados will be discussing some of the important issues concerning the current cell lines that are being used in baculovirology. In addition, he will also address future developments that may lead to novel cell lines that will be useful in biotechnology.

10:05 - 10:35 Coffee Break, Exhibit and Poster Viewing

BEVS--More than Making Proteins in Insect Cells

10:35 - 10:40 Chairperson's Remarks

 

10:40 - 11:10 KEYNOTE PRESENTATION

Baculovirus-Mediated Gene Transfer into Mammalian Cells  Frederick M. Boyce, M.D., Ph.D., Department of Neurology, Massachusetts General Hospital, Harvard Medical School Baculoviruses can be modified to transfer genes into mammalian cells in addition to their better-known role as vectors for protein expression in insect cells. Baculoviral vectors have a number of advantages over conventional animal viral vectors for gene transfer into mammalian cells, including ease of construction and use, large packaging size, and increased biosafety. The applications of baculoviruses for gene therapy, large-scale protein expression in mammalian cells, and high-throughput screening will be discussed.

11:10 – 11:40 Enhanced Gene Delivery by Modified Baculoviruses
Kari Airenne, Ph.D., Department of Biotechnology and Molecular Medicine, A.I. Virtanen Institute, University of Kuopio
Autographa californica multiple nucleopolyhedrovirus seems to have a capacity to enter all types of cells from different origin. Transgene expression is achieved not only in the natural host (insect) cells but also in non-target cells if a suitable promoter is used. This provides the basis for the universal expression concept in which one-step cloning allows the expression of a desired gene in different hosts by using a multi-promoter strategy. However, unsatisfactory expression rates are often achieved in cells other than cells of insect or hepatic origin. We are interested in resolving baculovirus transduction mechanisms and converting this knowledge into the construction of improved baculoviruses for recombinant protein production, drug screening, gene therapy, and other applications. The presentation will focus on the conceptual aspects of how to improve baculovirus technology. Some recent data of improved vectors showing substantial enhancement in gene delivery will also be presented.

11:40 – 12:15 Development of Baculovirus-Derived Transposon Insertions as Versatile Gene Transfer Vectors for Eukaryotic Organisms and Cell Cultures
Malcolm J. Fraser, Jr., Ph.D., Professor, Department of Biological Sciences, University of Notre Dame
The piggyBac transposon was first identified as one of several insertional mutations in cell culture passaged Baculoviruses. The helper-dependent trans-mobilizing capabilities of this transposon for gene vectoring was first established in the Baculovirus system. Later applications of this transposon to a diversity of organisms including protists, invertebrates, and vertebrates has established it as a highly versatile tool for genetic engineering. The unique properties of piggyBac mobilization provide novel capabilities including saturation gene tagging, promoter/enhancer trapping, relatively large carrying capacity, and precisie excision from insertion points within a genome. Several recent applications illustrative of the versatile capabilities of this transposon vector will be discussed.

12:15 - 1:45 Lunch on Your Own (Sponsorship Available)

1:45 – 1:50 Chairperson's Remarks

 

1:50 - 2:25 FEATURED PRESENTATION

Sweetening the Pot: Developing Engineered Baculovirus Expression Systems for Humanized Recombinant Glycoprotein Production Donald L. Jarvis, Ph.D., Professor, Department of Molecular Biology, University of Wyoming The baculovirus-insect system is widely used for recombinant protein production. This system can provide high-level production and, because insect cells are eukaryotic, it can provide eukaryotic protein modifications, such as glycosylation. On the other hand, insect protein glycosylation pathways are not identical to those of higher eukaryotes. Therefore, the baculovirus-insect cell system typically fails to produce recombinant glycoproteins with oligosaccharide side chains identical to those found on native, higher eukaryotic products. For the past decade, we have been addressing this problem by using genetic engineering of both the baculovirus vector and the insect cell host in an effort to humanize the protein N glycosylation pathway in this system. This presentation will focus on these efforts with an emphasis on transgenic insect cell lines with humanized N-glycosylation pathways.

Protein Production

2:25 - 2:55 Sorting Out the Variables of Protein Expression in BEVS
William Gillette, Ph.D., Senior Scientist & Leader, Protein Purification Group, Protein Expression Laboratory, SAIC-Frederick, Inc., NCI-Frederick
We have established a standard screen for determining the optimal conditions for expressing soluble protein in the BEVS. Each of the following factors can have a dramatic impact on expression: insect cell line, incubation temperature, harvest time, and fusion protein. We have shown that these factors affect expression levels, protein solubility, and the degree of target proteolysis. The effect of these variables on successful expression is target protein dependent and not currently predictable, although trends are evident and may be useful for improving the success rate of high throughput operations.

2:55 - 3:10 Increasing Recombinant Protein Expression in Insect Cells
Thera Mulvania, Ph.D., Expression Systems LLC
It is of great interest to increase the efficiency and productivity of recombinant protein expression for both small- and large-scale applications. Efforts to increase recombinant protein production can be made at the level of culturing conditions, cell line choice, construction of the vector/virus, and infection strategies. Here we will discuss the impact of varying these significant parameters of the Baculovirus expression system. 

3:10 – 3:25 The Development of Biological and Automated Tools to Improve Recombinant Baculovirus Production and Protein Expression
Geoff Alms, NextGen Sciences Inc.
The baculoworkstation™ increases throughput of recombinant baculovirus production by automating key methods (insect cell seeding, transfections, viral dilutions, and small-scale protein production). The baculoworkstation™ is compatible with a variety of commercially available baculovirus technologies for example, flashBAC™ a new one-step baculovirus production technology.

3:25 - 4:00 Achievements in Large-Scale Production of Gene Therapy Viral Vectors Exploiting the Baculovirus-Insect Cell System
Alejandro Negrete, Ph.D., Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health
The use of viral vectors in gene therapy clinical trials and commercialization require optimized large-scale production processes. For process optimization during scale-up studies, monitoring on-line conductivity and permittivity at different frequencies provide valuable information about baculovirus infection of Sf9 cells and recombinant adeno-associated virus production. It is possible to follow-up the infection, detect culture changes, and determine the optimal harvesting time in order to achieve higher yields.

4:00 – 4:30 Refreshment Break, Exhibit and Poster Viewing

4:30 – 5:00 Rapid and Scalable Production of Viral Antigens in the PERLXpress in vivo Baculovirus-Based Protein Production System
Dr. George Buchman, Chief Scientific Officer, Chesapeake PERL, Inc.
Chesapeake PERL is using its proprietary PERLXpressTM system to produce viral antigens for protein banking and for the development of subunit vaccines. The system enables the production of initial quantities of protein in as little as three weeks followed by the rapid, linear scale-up to gram and kilogram quantities. Data will be presented on a series of proteins derived from the vaccinia virus with potential application as a third generation smallpox vaccine. These antigens possess immunological properties indistinguishable from proteins produced in cell culture, including the induction of neutralizing antibodies in mice.

5:00 – 5:30 A Comparison of Recombinant Protein Production Using Baculovirus and Mammalian Transient Transfection Technologies
Christopher Kemp, Ph.D., Kemp Biotechnologies, Inc.
The baculovirus expression system is an excellent tool for the production of milligram to gram quantities of complex recombinant proteins. Recent advances in the technology of mammalian transient transfection provide a second option for the production of proteins historically produced in baculovirus-infected insect cells. This presentation will compare the production of various protein types in both systems.

5:30 – 6:00 MultiBac: New Baculovirus Expression Tools for Multiprotein Applications
Imre Berger, Ph.D., P.D., Group Leader, Institute for Molecular Biology and Biophysics, Swiss Federal Institute of Technology ETH
The concept of the cell as a collection of multisubunit protein machines, one each for essentially every major process, has become a cornerstone of modern biochemistry. Molecular level study of these complexes often depends on recombinant production for sufficient quantity and homogeneity of the sample. MultiBac is a simple and versatile system for generating baculovirus DNA to express protein complexes comprising many subunits. To facilitate the assembly of multigene expression casettes, this method uses transfer vectors containing a multiplication module that can be nested, as well as recombination elements to minimize time-intensive use of nucleases and ligase. The simplicity and remarkable robustness of the method are illustrated by production and characterization of several large multisubunit eukaryotic transcription factor complexes.

6:00 – 7:00 Reception in the Exhibit Hall

7:00 End of Day One 

For more information, please contact:
Mary Ruberry, Conference Producer
Phone: 781-972-5421 • E-mail: mruberry@healthtech.com  

For exhibit and sponsorship information, please contact:
Suzanne Carroll, Manager, Business Development
Phone: 781-972-5452 • E-mail: scarroll@healthtech.com