Antibody Engineering for Expression in Insect Cells and Larvae

Antibodies are currently deployed as the recognition component of sensors that detect biological threat agents. Previously, we developed an anti-botulinum toxin antibody using a bacterial host. However, antibodies expressed in bacteria lack molecular modifications made post-translationally by animal...

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Bibliographic Details
Main Authors: Anderson, Patricia E, Valdes, James J, O'Connell, Kevin P, van Beek, Nikolai A, Liu, Yi, Davis, David, Bentley, William, Chase, Terry E
Format: Report
Language:English
Subjects:
ANTIBODIES
BACTERIA
BIOLOGICAL AGENTS
BIOLOGICAL WARFARE
CELLS
CELLS(BIOLOGY)
CHAINS
CODING
DETECTORS
ENGINEERING
GENES
INSECTS
LARVAE
Medicine and Medical Research
MODIFICATION
MOLECULES
PLASMIDS
RECOGNITION
SYMPOSIA
THREATS
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Summary:Antibodies are currently deployed as the recognition component of sensors that detect biological threat agents. Previously, we developed an anti-botulinum toxin antibody using a bacterial host. However, antibodies expressed in bacteria lack molecular modifications made post-translationally by animal (eukaryotic) cells. It was therefore desirable to express Fab fragment antibody genes in insect cell lines and larvae. In this study, we improved an existing baculovirus expression vector by inserting the reporter gene DsRed, then modified and inserted the heavy and light chain genes encoding an anti-botulinum toxin-binding Fab antibody. The structures of all plasmids constructed were verified by restriction analysis and sequencing. Preliminary data demonstrate that DsRed and anti-botulinum Fab are both strongly expressed in larvae of Trichoplusia ni, suggesting that this system may be an economical manufacturing process for recombinant antibodies. See also ADM001523. The original document contains color images. Pub. in Proceedings of the Joint Service Scientific Conference on Chemical and Biological Defense Research (2002) Held in Hunt Valley, MD on 19-21 Nov 2002.