View All Protein Production at Instruct
The NKI Protein Facility is highly experienced in protein production using insect and mammalian cells, and assists users with the design of expression constructs considerinbgt also solubility and domain boundaries. We collected a wide variety of expression vectors, including LIC vectors for Ligand Independent Cloning. For expression of intracellular and extracellular proteins in insect cells we have SF9 and SF21 systems. Our mammalian expression system is optimised for HEK-293T cells, but we have for instance also the glycosylation free, gnT1-deficient HEK293S variety.
Access to protein expression in bacteria (E. coli) is possible with the NKI Proteikn Facility, but not accessible via Instruct!
Depending on the project, the platform for expression will be chosen. Proteins can be expressed in E. coli, Insect cells and mammalian cells. For each of the systems we can do small scale (few ml) as well as large scale (few liters) production. For insect cell expression SF9 and SF21 cells are used for expression of intracellular and extracellular proteins. The cells are grown in suspension cultures in volumes up to 500 ml/flask. We use Baculovirus technology for over-expression of proteins in insect cells.
Depending on the project, the platform for expression will be chosen. Proteins can be expressed in E. coli, Insect cells and mammalian cells. For each of the systems we can do small scale (few ml) as well as large scale (few liters) production. Our mammalian expression system has currently been optimized for HEK-293T cells and is typically used for expression of extracellular (secreted) proteins. These cells can be cultured in flasks, dishes or rollerbottles. A glycosylation deficient strain - gnT1-deficient HEK293S - is also available for production of eukaryotic proteins that would benefit from removal of glycosyl chains (e.g. for crystallisation purposes). The use of other cells for expression can be discussed.
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