Antibody Engineering for Expression in Insect Cells and Larvae
Abstract
Antibodies are currently deployed as the recognition component of sensors that detect biological threat agents. Antibodies that detect simulants of bio-threat agents are also currently incorporated into detection platforms for testing and evaluating new devices and materials. Previously, we developed an anti-botulinum toxin antibody using a powerful genetic technology known as phage display, in which a very large library of immunoglobulin (antibody) genes are expressed on the surface of bacteriophage (bacterial virus) particles. 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. The use of insects as gene expression "bioreactors" poses far fewer ethical concerns than the use of mammalian systems. 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 the reporter gene DsRed is strongly expressed in larvae of Trichoplusia ni, suggesting that this system may be an economical manufacturing process for recombinant antibodies.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2002
- Accession Number
- ADA405911
Entities
People
- James J. Valdes
- Kevin P. O'connell
- Patricia E. Anderson
Organizations
- Edgewood Chemical Biological Center