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Insect Cell Culture and Baculovirus Expression System
Insect Cell Culture and Baculovirus Expression System
Our research focuses on the culture of Spodoptera frugiperda (Sf) cells and the production of recombinant proteins using the baculovirus expression vector system (BEVS).
Insect cell lines, including the Sf9 line derived from S. frugiperda ovarian tissue, have become a well-established platform for recombinant protein production at both laboratory and industrial scale. These cells combine ease of handling with the capacity to express complex, post-translationally modified eukaryotic proteins. The BEVS exploits the high infectivity and genetic capacity of baculoviruses, specifically Autographa californica multiple nucleopolyhedrovirus (AcMNPV), to introduce and express foreign genes in insect cells. The system enables high-level expression under the control of strong viral promoters and supports the production of a wide range of protein types. Due to its versatility, scalability, and ability to support co-expression of multiple proteins, the BEVS is widely used for the generation of therapeutic proteins, vaccine candidates, viral vectors, and structural biology targets.
Here you will find the focus of current and past research in our group.
Improving Insect Cell Cultivation
We are investigating strategies to improve the efficiency and reproducibility of Sf9 cell culture. One focus is the modification of the surface properties of glass culture vessels. We have shown that silanization has positive effects on cell growth, viability and experimental reproducibility by minimizing unwanted cell adhesion and aggregation.
Establishing different methods for quantification of virus titer
The multiplicity of infection (MOI) is critical to infection progression and product yield. A low MOI can lead to incomplete infection, whereas a high MOI can lead to premature cell death. Accurate quantification of the virus concentration is therefore essential to calculate the MOI. In this context, we have established an mCherry- and GP64-based qPCR, a reliable plaque assay and a cell size-based assay.
Optimization of transfection and infection of Sf9 cells
Efficient transfection and infection are crucial for the production of recombinant proteins using the BEVS. Targeted optimization of these processes - in both adherent and suspension cultures - can significantly improve virus yield, protein quality and yield, reproducibility and scalability. For example, defined virus addition (MOI) and the correct harvest time can minimize cell stress, promote synchronous expression and increase protein expression. Especially in large-volume suspension systems, precise infection control is essential for the successful use of the system in research and industry.
Production and functional characterization of an integral transport protein from the SLC2 family
As part of a doctoral thesis, a membrane transport protein is to be produced and functionally characterized using BEVS. Transport proteins are crucial for the uptake and distribution of molecules in cells and organisms. The establishment of a stable platform to produce these difficult-to-express proteins enables the analysis of their biological properties and functionality and helps to identify potential biotechnological and medical applications.
Upscaling baculovirus-mediated protein production
Upscaling cultivation and protein production from laboratory to reactor scale enables the expression of large amounts of target protein. One challenge is the formation of foam, which negatively affects cell growth, protein yield and process control. We have shown that antifoam agents have a positive effect on Sf9 cell growth and can also increase protein production.
Generation of an mCherry reporter system
For the microscopic monitoring of baculovirus-mediated protein expression, a reporter system with the fluorescent mCherry was established. Several constructs have been developed for the co-expression of mCherry with target proteins as well as for the expression of mCherry fusion proteins. The system has already been successfully applied to the fusion of mCherry with a membrane-bound protein, allowing the targeted study of localization and expression in insect cell cultures.
Completed theses and student research projects