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Production of Recombinant PRMT Proteins using the Baculovirus Expression Vector System
Published on: July 17, 2021
Generation of lentivirus vectors using recombinant baculoviruses
H P Lesch1, S Turpeinen, E A Niskanen
1Department of Biotechnology and Molecular Medicine, AI Virtanen Institute for Molecular Sciences, University of Kuopio, Kuopio, Finland.
Gene Therapy
|May 9, 2008
Summary
Baculovirus technology provides a scalable method for producing lentiviral vectors in mammalian cells, achieving titers comparable to traditional methods without generating replication-competent lentiviruses.
Area of Science:
- Biotechnology
- Virology
- Molecular Biology
Background:
- Clinical-scale lentiviral vector production faces challenges with current transfection and stable cell line methods.
- Baculovirus technology offers advantages in scalability, speed, mammalian cell transduction efficiency, and safety.
Purpose of the Study:
- To evaluate baculovirus technology as a scalable system for producing replication-defective lentiviral vectors.
- To construct recombinant baculoviruses encoding essential lentiviral vector components.
Main Methods:
- Four recombinant baculoviruses were constructed: BAC-transfer (GFP transgene), BAC-gag-pol, BAC-VSVG, and BAC-rev.
- 293T cells were transduced with recombinant baculoviruses to produce lentiviruses.
- Optimization of baculovirus concentration, medium, and transduction time was performed.
Main Results:
- Functional lentiviruses were produced with unconcentrated titers averaging 2.5 x 10^6 TU/ml, comparable to conventional methods.
- Produced lentiviruses successfully transduced HeLa cells, demonstrating sustained GFP expression.
- p24 ELISA confirmed the absence of replication-competent lentiviruses.
Conclusions:
- Baculovirus-mediated production is a viable and attractive alternative for scalable lentiviral vector generation in mammalian cells.
- This system demonstrates safety and efficiency, addressing limitations of current lentiviral vector production methods.
