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Protein Expression Platforms

 

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.

 

E. coli
  • A set of different expression strains is available from the facility, including BL21 (DE3) (+/- pLysS) and Rosetta2 (DE3) (+/-pLysS). These strains have been adopted by the facility and are all T1-phage resistant. No other strains are allowed to be used within the facility.

  • Expression trials can be performed in volumes as low as 2 ml (in 24-well format). Incubators are available for large scale expression (up to 6 Liter/ incubator) at variable temperatures.

Insect cells
  • 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 technolgy for over-expression of proteins in insect cells.

 

Mammalian cells
  • Our mammalian expression system has currently been optimized for  HEK-293T cells. 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.

NKI Protein Facility                                                                                                                                                                                                         Last updated: 13 April 2012

NKI-AvL

Plesmanlaan 121

1066 CX Amsterdam

The Netherlands