Available Services

The service offers dedicated pipelines for small- and large-scale cell cultures and expressions:

• Small scale: 15 litres fermenter useful for initial trial test before moving to larger scale fermenter or small expression using both bacteria and yeast cells. Typical culturing strategies are available such as batch and fed batch (up to 2 feeds). Max working volume of 10 litres.
• Large scale: 150 litres fermenter dedicated to large cultivation and protein production. Typical cultivation strategies are available, including batch and fed batch (up to 2 feeds). Max working volume of 100 litres.

In both cases, cells will be collected and separated from the supernatant using dedicated superspeed centrifuges or a High-Speed Tubular Centrifuge. Upon request, the cells can be lysed using a continuous flow cell disruptor or by a high-capacity cryogenic grinder.

Go to the open call

The service offers dedicated pipelines for small- and large-scale cell cultivation and protein expressions:

• Small scale: utilizing commercial Expi293F cells, transient transfection is conducted with a plasmid containing the gene\s of interest (potentially with a fluorescent marker), provided by the applicant. Cell cultivation and transfection take place in Erlenmeyer flasks within dedicated shaker incubators; typically, 500 ml of cell culture per 2-liter flask is utilized up to a total cultivation volume of approximatively 10 litres. Transfection of an Expi293F cell culture is performed at approximately 2 to 3 x10^6 cells\ml, following a modified protocol with polyethylenimine (PEI). While no expression test will be performed, the efficiency of transfection can be readily assessed if the plasmid includes a fluorescent marker. Transfected cells are subsequently harvested by centrifugation, and either the frozen pellet or the refrigerated supernatant will be provided.
• Large scale: bioreactor cultivation and protein expression are performed either with commercial cell lines (e.g., Expi293F cells) transiently transfected with applicant-provided plasmids, or with any mammalian cell line adapted to suspension cultivation supplied by the applicant. Preliminary tests are normally performed using a 0.5 litres scale bioreactor to define and optimize bioreactor cultivation conditions. Subsequently, a 10 litres scale bioreactor enables large-scale cell cultivation and protein expression, employing both transient transfection protocols and stable cell line expressions. Moreover, the implementation of perfusion, utilizing an acoustic retention device, may be explored to increase the biomass production and, consequently, protein expression. Similar to small-scale operations, no expression test will be performed. Instead, monitoring of transfection efficiency for transient expression can be undertaken if the plasmid carries a fluorescent marker or fluorescently tagged protein of interest.

Go to the open call

The service aims to cover the entire process of Baculovirus protein expression, starting from a Bacmid. Initially, commercial Sf9 cells are transfected with the Bacmid, supplied by the applicant, which contains the gene of interest and a fluorescence marker. The initial stage results in a low-titre Baculovirus suspension (P1 generation), followed by a second amplification (P2 generation) to produce a high-titre suspension. The P2 Baculovirus suspension could be directly used for protein expression or another round of Baculovirus amplification is performed before infecting commercial High Five insect cells (at 0.5×10^6 cells\ml). The virus amount used depends on the construct and might need optimization in initial small-scale infections. Protein expression is monitored indirectly by checking for the fluorescence marker (carried by the Bacmid) and cell viability. Finally, cells are harvested via centrifugation, and the pellet or supernatant is made available.

Go to the open call