General information
The Baculovirus insect cell expression system is widely used for the production of precisely matured, folded and processed recombinant proteins. In contrast to a prokaryotic expression system like E. coli, insect cells are able to glycosylate the proteins although the glycosylation pattern is not identical to mammalian cells. This unique tool usually yields high amounts of the produced protein, making it highly cost effective in comparison to other eukaryotic expression
systems.
Test Phase (please refer to Table 1)
- Cotransfection of insect cells with transfer vector DNA and linear virus DNA,
generation of recombinant viruses, plaque purification
- Test expression of 6 recombinant viruses
- Western blot analysis
- Estimation of the required culture volume to obtain the requested amount
of protein
- After having performed the test expression the customer receives
supernatants (2 ml) or cell pellets of 6 test expression cultures for his own
tests
- Time required: 4-8 weeks
Production Phase (please refer to Table 1)
- High titer virus stock production
In case of detectable and functional recombinant protein in at least one of the 6 samples, the recombinant baculovirus has to be amplified to get a high titer virus stock (100 ml of 5 x 107 to 1 x 108 pfu/ml)
- Time required: 1-3 weeks
- Culture in the appropriate scale and protein expression
- Cell harvest and lysate preparation
- Ni-NTA affinity purification
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