The host cell line could be infected with recombinant virus after a certain concentration of virus was obtained.
In theory, insect cells will begin to express the target protein about 20-24 after being infected by recombinant baculovirus. In reality, however, the exact expression timing is always elusive. According to the characteristics of baculovirus and insect cells, protein expression can be divided into four stages, early, middle, late, very late stage. The target gene sequence is usually inserted downstream of the extremely late overexpression promoter Polh and P10. Thus, the expression of the target protein will not start until about 20-24 hours after infection. However, for different proteins and different expression patterns (intracellular and secretory), the time for protein expression to start or reach the peak is not certain. Researchers not only need to distinguish the early expression protein and the target protein but also need to monitor the protein expression curve in order to harvest the target protein at the peak value. Missing the peak protein expression time means that the yield and quality of the target protein are decreased.
As an expert in protein expression, Creative BioMart is always willing to help our customers. We provide high quality virus infection services, protein expression monitoring services, protein harvesting and processing services. At the same time, we recommend that you continue to choose our protein purification, qualitative and quantitative assay services after obtaining the target protein. Of course, if you choose to end the service, we will deliver the protein expression products to you as soon as possible.
Infect the Host Insect Cells with Recombinant Virus
MOI of 0.50, 1.0, 5.0, and 10.0 were carried out to determine the best optimal infection MOI (please refer to our Virus Amplification Service for details)
Optimizing Heterologous Protein Production
The BEVS recombinant gene product may or may not be secret. The peak expression of secretory protein usually appeared 30-72 hours after infection while that of non-secretory proteins usually reached the peak at 48-96 hours post-infection. To be on the safe side, after 24 hours post-infection, samples will be taken every 8-12 hours to determine the protein expression profile and the optimal harvest time.
| Insect Cell | Advantage | Use |
| Sf9 (Spodoptera frugiperda) | Well-characterized host cell, good for plaquing | All general types of recombinant baculovirus production and protein expression |
| Sf21 (Spodoptera frugiperda) | High yield of intracellular protein | Intracellular recombinant baculovirus production and protein expression |
| High Five (Trichoplusia ni) | High yield of secreted protein, shorter culture period | Secreted protein expression |
| BmN (Bombyx mori L.) | High yield cell line | For BmNPV and HyNPV expression platform |
If you are interested or have questions about our service, please contact us.
Creative BioMart has been committed to protein expression, purification and analysis for many years, and has established an advanced technology platform. Our professional technical team can provide appropriate protein sequence services according to the protein provided by customers.